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Stem cells can differentiate to diverse cell types in our body, and they hold great promises in both basic research and clinical therapies. For specific stem cell types, distinctive nutritional and signaling components are required to maintain the proliferation capacity and differentiation potential in cell culture. Various vitamins play essential roles in stem cell culture to modulate cell survival, proliferation and differentiation. Besides their common nutritional functions, specific vitamins are recently shown to modulate signal transduction and epigenetics. In this article, we will first review classical vitamin functions in both somatic and stem cell cultures. We will then focus on how stem cells could be modulated by vitamins beyond their nutritional roles. We believe that a better understanding of vitamin functions will significantly benefit stem cell research, and help realize their potentials in regenerative medicine.
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Cellular and Molecular Life Sciences
Roles ofvitamins instem cells
CarlosGodoy‑Parejo1· ChunhaoDeng1· YumengZhang1· WeiweiLiu1,2· GuokaiChen1,2,3
Received: 22 July 2019 / Revised: 12 October 2019 / Accepted: 21 October 2019
© Springer Nature Switzerland AG 2019
Stem cells can differentiate to diverse cell types in our body, and they hold great promises in both basic research and clinical
therapies. For specific stem cell types, distinctive nutritional and signaling components are required to maintain the pro-
liferation capacity and differentiation potential in cell culture. Various vitamins play essential roles in stem cell culture to
modulate cell survival, proliferation and differentiation. Besides their common nutritional functions, specific vitamins are
recently shown to modulate signal transduction and epigenetics. In this article, we will first review classical vitamin functions
in both somatic and stem cell cultures. We will then focus on how stem cells could be modulated by vitamins beyond their
nutritional roles. We believe that a better understanding of vitamin functions will significantly benefit stem cell research,
and help realize their potentials in regenerative medicine.
Keywords Vitamin· Cell culture· Vitamin A· Vitamin B3· Vitamin C· Vitamin E· Embryogenesis· Stem cells
Vitamins are natural organic compounds that play essen-
tial roles in normal physiological functions in minimum
amounts, but the host either cannot synthesize them, or can-
not produce an adequate amount to meet the normal physi-
ological demands [1]. The word vitamin comes from the
Latin word “vita” meaning “life”, which reflects its essential
roles in the survival and well-being of humans [2]. Vitamins
are involved in diverse cellular functions, and their defi-
ciency often leads to serious symptoms to people, sometimes
even death [3]. Since the discovery of vitamin A in 1912,
13 vitamins have been identified based on their essential
roles in human health [4]. Most vitamins can be obtained
through balanced food intake, and vitamin supplements are
also widely used in healthcare practices. In the 1950s, peo-
ple found that vitamin supplements are also essential for
invitro cell culture due to their nutritional functions [5, 6].
Recently, various vitamins are shown to possess regulatory
mechanisms on the cellular level, especially in stem cells [7].
Stem cells are a special group of cells that can proliferate
extensively and have the potential to generate various cell
types in the human body [8]. Embryonic stem cells (ESCs)
are pluripotent and can differentiate to all cell types. ESCs
only transiently exist during embryogenesis, and finally give
rise to all the cells in an embryo. Adult stem cells possess
limited potential to differentiate to specific cell types, and
can be classified into multipotent and unipotent stem cells
[9]. They are responsible for the daily maintenance and
repair of tissues [10]. With somatic reprogramming tech-
nologies, stem cells can now be generated from somatic cells
with defined factors [11]. Stem cells are widely used in basic
research to understand embryogenesis and homeostasis, to
model diseases, and are also important source materials for
cell therapies in regenerative medicine [12]. Most stem cell-
related studies and applications involve cell culture systems,
which provide essential components for specific cell types to
survive and properly exert their normal functions.
A typical cell culture system normally contains ten cat-
egories of components, including water, inorganic salts,
growth factors, amino acids, buffering reagent, energetic
substrates, extracellular matrix, vitamins, vitamin-like
organic factors and the cell culture atmosphere. Functional
stem cells require a culture system in which all components
Cellular andMolecular Life Sciences
* Guokai Chen
1 Centre ofReproduction, Development andAging,
Faculty ofHealth Sciences, University ofMacau, Taipa,
MacauSAR, China
2 Bioimaging andStem Cell Core Facility, Faculty ofHealth
Sciences, University ofMacau, Taipa, MacauSAR, China
3 Institute ofTranslational Medicine, Faculty ofHealth
Sciences, University ofMacau, Taipa, MacauSAR, China
C.Godoy-Parejo et al.
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are suitably balanced. To realize the great potentials of
stem cells in regenerative medicine, people often modulate
and optimize cell culture components to improve stem cell
functions. Regulation of signal transduction pathways with
growth factors has traditionally been the main approach [13,
14]. However, nutritional regulation is emerging as a viable
target for stem cell modulation, which could affect not only
cell survival but also pluripotency and cell fates [1517].
As an essential part of cell culture, the important roles of
vitamins are manifested in our daily use of cell culture in
basic research and clinical applications. This article will try
to review how vitamins are utilized in stem cell applications.
We will first introduce the general vitamin requirements in
cell culture. Then we will focus on vitamin A, vitamin B3,
vitamin C and vitamin E, and discuss how they are utilized
in stem cell applications [1821].
A brief background onvitamins
inthehuman body
Human vitamins are generally categorized into two classes,
nine water-soluble vitamins and four fat-soluble vitamins
(Table1) [22]. Water-soluble vitamins include 8 members of
the B type vitamins and vitamin C, and fat-soluble vitamins
include vitamins A, D, E and K. All the vitamins can be
obtained from food to fulfill the nutritional needs (Table1).
Some vitamins can be synthesized in the human body, but
at a very low rate (Table2) [23, 24]. In this review, we will
briefly summarize some key vitamin-dependent processes
and the role these vitamins play in stem cell biology.
All the water-soluble vitamins are coenzymes for impor-
tant metabolic enzymes that are essential for cellular
functions. Their essential roles in metabolic pathways are
illustrated in Fig.1. Vitamins B1, B3, B6 and B7 are involved
in glucose metabolism that includes glycolysis, pentose
pathway, glycogenolysis and gluconeogenesis. Fatty acid
synthesis and degradation require vitamins B2, B3 and B5.
Meanwhile, amino acid degradation requires vitamins B3,
B6, B9 and B12. The TCA cycle and oxidative phosphoryla-
tion take place in mitochondria, and utilize vitamins B1, B2,
B3, B5 and B7 in specific steps. Often times, multiple vita-
mins are involved in the same metabolic process. For exam-
ple, When acetyl-CoA is generated from pyruvate by pyru-
vate dehydrogenase, four of the five coenzymes involved
in this step are vitamins, including vitamins B1, B2, B3 and
B5 [25, 26]. Any deficiency in these vitamins could lead to
malfunction of the TCA cycle.
Besides type B vitamins, other vitamins’ functions are
more diverse. Vitamin C is the only water-soluble vitamin
that does not belong to the vitamin B family, and it is known
to regulate collagen synthesis by acting as a cofactor for pro-
lyl hydroxylases, reducing its iron center [2729]. In addi-
tion, vitamin C is an antioxidant that suppresses the produc-
tion of reactive oxygen species (ROS). It is well known for
its role in the prevention of scurvy [30, 31]. Vitamin A fam-
ily members have distinctive functions, including the preven-
tion of night blindness. At the molecular level, vitamin A
functions through antioxidation and transcriptional regula-
tion [32, 33]. Vitamin D is a hormone that binds to nuclear
receptors to regulate transcription, and it is best known for
its role in calcium absorption [34]. Vitamin E is a potent
fat-soluble antioxidant. Some vitamin E isoforms were also
reported to modulate signal transduction [35]. Vitamin K
is a cofactor for γ-glutamyl carboxylase that is essential for
blood clotting [36, 37].
Table 1 Vitamins and their functions [25, 26, 34, 36, 71, 92, 161, 166, 199, 207, 248, 249, 251, 253259]
Vitamin Names Daily dose Cellular function
Vitamin B1Thiamine 1.2mg Glycolysis, non-oxidative phase of pentose pathway
Vitamin B2Riboflavin 1.2mg Coenzyme in carbohydrate and lipid metabolism; activation of B6 and
B9; antioxidant
Vitamin B3Niacin, nicotinamide, nicotinamide riboside 15mg Coenzyme in carbohydrate, amino acid and lipid metabolism
Vitamin B5Pantothenic acid 5mg Coenzyme in carbohydrate and lipid metabolism, lipid biosynthesis
Vitamin B6Pyridoxine, pyridoxamine, pyridoxal 1.5mg Coenzyme in glycogenolysis and amino acid metabolism
Vitamin B7Biotin 30µg Lipid synthesis; leucine catabolism; conversion of amino acids and
propionate to glucose in liver; gluconeogenesis
Vitamin B9Folic Acid 400µg Coenzyme in nucleotide synthesis, methylation of chromatin, DNA,
RNA, histone and transcription factors, amino acid metabolism
Vitamin B12 Cobalamin 2.4µg Coenzyme in folate and homocysteine metabolism
Vitamin C Ascorbic acid 85mg Antioxidant; coenzyme in collagen synthesis
Vitamin A Retinoic acid, retinol, all-trans-RA 800µg Vision, cell differentiation, reproduction
Vitamin D Cholecalciferol, ergocalciferol 15µg Mg, Ca and P absorption
Vitamin E Tocopherols, tocotrienols 15mg Antioxidant, cell membrane integrity
Vitamin K Phylloquinones, menaquinones 115µg Protein synthesis in blood coagulation
Roles ofvitamins instem cells
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Essential vitamins inregular cell culture
Because of vitamins’ important functions, they are essen-
tial not only for the whole organism but also for individual
cells. However, the vitamin dependency of the human body
is often different from cells in culture media. The impor-
tance of individual vitamins is gradually discovered through
the years. In 1950, Morgan and colleagues first showed that
cell survival was improved by a vitamin mixture in serum-
free synthetic medium [38]. In 1955, Eagle systematically
analyzed the impact of individual vitamins on the growth
of both mouse fibroblasts and Hela cells [39]. Six vita-
mins were shown essential for cell proliferation of both cell
lines. They all belong to the B vitamin complex, including
B1, B2, B3, B5, B6 and B9. The medium was named Basal
Medium Eagle (BME), which is the first synthetic medium
with defined vitamin functions. In the following years, Mini-
mum Essential Medium (MEM) and Dulbecco’s Modified
MEM (DMEM) were developed with increased amino acid
or vitamin concentrations, but the vitamin composition still
remained at six [40, 41].
Although the above basic media can support short-term
proliferation of a few cell lines, their capacity is insufficient
for many other lines in long-term culture. To support clonal
growth and long-term culture of Chinese hamster cell lines,
Ham developed the Ham’s F-10 and F-12 media that contain
additional B7 and B12 [42, 43]. B7 was important for the cell
growth and viability of a variety of cell types [44], while
vitamin B12 was found to be essential for lipid metabolism
[45, 46]. People later found that cell growth is improved
when DMEM and Ham’s F12 are mixed in a 1:1 ratio, and
this medium was named DMEM/F12 [47]. The eight B vita-
mins (B1, B2, B3, B5, B6, B7, B9 and B12) in DMEM/F12 are
also present in a variety of other basic media such as RMPI,
IMDM and α-MEM [48]. These vitamins are generally con-
sidered as essential vitamins for most cells cultured invitro.
DMEM/F12 is the most commonly used base medium for
human embryonic stem cells [17, 4951], so we will use
Table 2 Vitamins in cell culture media [1, 39, 44, 45, 47, 50, 71, 79, 155, 157, 235, 260265]
Vitamin Concentration in blood Concentration
in DMEM/F12 Endogenous source Application in stem cells
Vitamin B166–200nM 6.44µM Gut bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B2174–471nM 0.58µM Colon bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B381–213nM 16.6µM Biosynthesis from tryptophan
Colon bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B50.5–1.9µM 4.7µM Colon bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B615–73nM 9.8µM Colon bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B7> 400ng/L
1.64nM 14.3nM Hindgut bacteria In most base medium, such as BME,
α-MEM, Ham’s F12 and DMEM/F12;
Used for somatic and stem cell culture
Vitamin B9> 3.0nM 6µM Gut bacteria In all the base medium; used for somatic
and stem cell culture
Vitamin B12 118–716pM 50nM Gut bacteria; not clear whether B12 can
across colon In most base medium, such as α-MEM,
Ham’s F12 and DMEM/F12; Used for
somatic and stem cell culture
Vitamin A 1.4–3.2µM Retinol promotes self-renewal and pluri-
potency; Retinoic acid mainly drives cell
differentiation by modulating epigenetics
Vitamin C 25–85µM In kidney and liver (except high primates) Supports cell reprogramming, survival and
collagen production; Reduces ROS
Vitamin D 30–100μg/L Precursor synthesized in the sebaceous
glands of the skin Leukocyte production and differentiation
Vitamin E 20–35µM Component of B-27 supplement and
chemically defined lipid concentrate;
Protects stem cells and progenitor cells
against oxidative stress; Affects ESC dif-
ferentiation through ROS levels
Vitamin K 0.22–2.22nM Menaquinones synthesized by bacteria in
the large intestine Promotes the differentiation of dental pulp
stem cells (DPSCs) to osteoblast invitro
C.Godoy-Parejo et al.
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DMEM/F12 as a reference to discuss vitamin formula and
concentration effect in this review.
When comparing vitamin composition in blood and in
DMEM/F12, there are two obvious discrepancies (Table2).
First, all B vitamins are present in DMEM/F12, but not other
vitamins including vitamins A, C, D, E and K. Consider-
ing all B vitamins are coenzymes for essential metabolic
processes, it is understandable that they are required for cell
culture. It also implies that the non-B vitamins are probably
not required for most cell types. It is also possible that those
vitamins could be provided through serum or medium sup-
plements such as B27. Second, all individual vitamins are
provided at significantly higher concentrations in DMEM/
F12 than in blood (Table2). It indicates that cells in culture
have differential reliance on vitamins.
Vitamin‑like nutrients forcell culture
In addition to essential vitamins, DMEM/F12 and many
other basic media also contain minute amount of some
organic compounds that are required to be supplemented
to the organism from food sources (Table3). We will brief
some of them here.
Choline can be biosynthesized from serine [52], and
it was first demonstrated as essential for cell survival and
proliferation in Eagle’s original vitamin study [39]. Cho-
line is essential for the generation of phosphatidylcholine
(PC) that is crucial for lipid transport and plasma membrane
integrity. Choline is also used to generate acetylcholine that
is important for neurotransmission [39, 53]. Choline can
serve as methyl donor in one-carbon transfer pathways, and
Fatty Acyl-CoA
B1, B2,
B3, B5
B2, B3, B5
Amino Acid Degradation
B3, B6
Fatty Acid Synthesis
Fatty Acyl-
Fatty Acid Degradation
Electron Transport Chain
Oxidative Phosphorylation
B2, B3
B1, B3, B5
Fatty acid
Amino acid
B2, B9
protein, lipid
One Carbon Metabolism
Fig. 1 Vitamin B in metabolism. B vitamins are essential for the major metabolic pathways. Specific vitamins (orange fonts) are highlighted in
multiple metabolic processes (blue fonts). OAA, oxaloacetic acid
Roles ofvitamins instem cells
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contribute to DNA modulation and histone epigenetic modi-
fication with the help of vitamins B9 and B12 [54].
Inositol can be naturally synthesized by the human body
from glucose in many tissues [55, 56], and Myo-inositol
was also identified by Eagle as an essential factor for cell
survival and proliferation in a wide variety of human cells,
both malignant and nonmalignant [57]. Myo-inositol is the
main source of phosphatidylinositol that mediates cell sig-
nal transduction, neurotransmission and osmoregulation [58,
Besides choline and inositol, essential fatty acids, such
as omega (ω)-3 and 6, are often found in culture media.
They are metabolized to form eicosanoids that affect lipid
homeostatic processes as well as the inflammatory response
[6063]. These lipids usually bind to albumin, and can be
supplemented to cells through albumin without notice.
Vitamin dependence incell culture
Cells in culture, including somatic and stem cells, have dif-
ferent vitamin dependency in comparison with the human
body as a whole. We believe that such difference is caused
by the inherent difference between the human body and arti-
ficial cell culture systems. First, not all vitamins that are
needed for the human body will be essential for cell cul-
ture. The deficiency of some vitamins often affects just one
or a few specific organs in the human body. For example,
vitamin K deficiency usually affects blood clotting but no
other physiological functions [37, 64]. When it comes to
cell culture, a vitamin may not be required for general cell
culture if it is needed for the survival and proliferation of
a specific cell type. Second, the human body usually has
specific organs to produce and store vitamins, which allows
people to tolerate temporary vitamin deficiency. However,
there is no endogenous backup mechanisms to complement
vitamin needs in cell culture, and all essential vitamins have
to be provided. If an essential vitamin is not provided in cul-
ture, severe symptoms often emerge quickly in cells. For this
reason, cell culture platforms have led to novel discoveries
of vitamin functions in recent years. Third, some nutrients
are not essential for the body, because specific organs can
produce sufficient amounts for all the cells in the body. How-
ever, in cell culture, these nutrients are considered vitamin-
like for cell culture, because they have to be provided for
normal cellular functions in the medium. Fourth, cell culture
is an artificial system, and nutrient concentrations in cell
culture can be modulated as needed. Often times, nutrients
can be tested and studied at concentrations that do not exist
in physiological conditions. Some novel vitamin-dependent
phenomena could only be identified in cell culture, in arti-
ficial conditions.
Differential vitamin dependence exists not only between
individual cells and the whole organism, but also among
different cell types. Vitamins affect metabolism similarly
in both somatic and stem cells, but they could have addi-
tional impacts on stem cells. In somatic cells, modulation
of specific vitamins will not change the cell identity. How-
ever, such changes might lead to loss of stemness or cell fate
changes in stem cells. A few vitamins have gathered inten-
sive interest in stem cell applications, and we will discuss
them in more details here.
Vitamin A
Vitamin A was the first vitamin discovered, and is actually
a group of compounds also known as retinoids, including
retinol, retinal and retinoic acid (RA) (Fig.2). Vitamin A
compounds are usually found in food of animal origin, while
their precursor, carotenoid, is present in plants. Humans can
synthesize vitamin A from carotenoids such as β-carotenes,
a lipid-soluble pigment responsible for the vivid colors in
plants. β-Carotenes can be converted into two retinals by
β-carotene 15,15-deoxygenase [65]. Retinal is then reduced
to retinol by retinaldehyde reductase, using NAPDH (vita-
min B3) as a cofactor. Retinol either is esterified by acyl-
transferases LRAT (lecithin-retinol acyltransferase) and
ARAT (retinol acetyltransferase) into retinyl palmitate for
storage, or is oxidized into retinoic acid by aldehyde dehy-
drogenase (ALDH) [66]. In human cells, retinal and retinol
are interconvertible; however, the conversion to retinoic acid
is irreversible [67].
Table 3 Vitamin-like factors [1, 266268]
Names Solubility Function Endogenous source
Choline Hydrophilic Lipid transport and metabolism, neuro-
transmission, methyl group donor Choline de novo synthesized through S-adenosylmethionine (SAM)-
dependent methylation of phosphatidylethanolamine by phosphatidyle-
thanolamine N-methyltransferase (PEMT). This process occurs mostly in
Hydrophilic Signal transduction and osmoregulation Inositol can be de novo synthesized from glucose, and the biosynthesis
occurs in brain, liver, and kidney
C.Godoy-Parejo et al.
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Although belonging to the same family, retinal, retinol
and retinoic acid play quite different roles in the human
body. For example, retinal is incorporated into the light sen-
sitive receptor rhodopsin in the retina, and prevents night
blindness. In contrast, retinoic acid can cause night blindness
by suppressing retinal production through the transcriptional
inhibition of ocular retinol dehydrogenases [33].
Vitamin A family members also play distinctive roles in
embryogenesis and stem cells in culture. Retinol and retinal
are readily oxidized in culture, so they can act as antioxi-
dants to promote cell survival and growth [32, 68]. Retinol
has been reported to help maintain the pluripotency and self-
renewal of hESCs [69], mESCs [70] and other progenitor
cells [7173]. In contrast, retinoic acid is a strong cell fate
modulator, which will be discussed in more detail below.
As a lipid-soluble compound, retinoic acid can diffuse
into the cytoplasm, bind to its nuclear receptor, and initi-
ate nuclear translocation and downstream regulation. Reti-
noic acid initiates the dimerization of retinoic acid receptor
(RAR) and retinoid X receptor (RXR). The heterodimer
then either directly regulates gene expression through a
DNA response element, or indirectly modulates transcrip-
tion through intermediate transcription factors [74]. Over
500 genes are influenced by the action of retinoic acid, and
many of the genes are involved in stem cell differentiation
and metabolism [74]. It was shown to be an inducer that
β-carotene 15,15’-deoxygenase
retinaldehyde reductase
Embryonic Asym metry/
Cell Proliferation
Stem Cell
Stem Cell
Ectoderm Inducer/
Cell Cycle Halter
Mesoderm/Endoderm Differentiation/
Maintenance and Viability of Progenitors
all-trans-Retinoic Acid
aldehyde dehydrogenase
Epigenetic Modulation/
Fig. 2 Vitamin A metabolism and function. Vitamin A is a group of
compounds derived from β-carotene. In humans, its alcohol isoform
has been reported to be beneficial for visual health. It also acts as an
antioxidant and promotes the pluripotency of stem cells. The acid
form, all-trans-retinoic acid, can interfere with the effect of retinol
in vision. Retinoic acid is a determinant for embryonic development,
and promotes cell proliferation. It can also promote reprogramming
of stem cells and differentiation of progenitors and act as a tumor sup-
Roles ofvitamins instem cells
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initiates differentiation in ESCs, and also a modulator in
lineage specific differentiation [75].
Retinoic acid modulates stem cell pluripotency and differ-
entiation through the expression of mRNA and microRNA
[21, 76]. It alters the expression of genes involved in DNA
methylation, histone acetylation and histone methylation. In
hESCs, the average level of DNA methylation is increased
by RA, promoting stem cell differentiation [77]. RA also
affect histone modifications, including acetylation of H3,
H4 and H3K in hESCs and mESCs, which leads to stem
cell differentiation [78, 79]. RA suppresses methylation in
H3K27 while promoting methylation in H3K4 in mESCs
and neuroblastoma, both stimulating cell differentiation [78].
At the same time, retinoic acid targets genes in metabolism,
cell proliferation and pluripotency. It usually suppresses
pluripotency gene expression, and promotes ectodermal dif-
ferentiation in ESCs upon the exit of self-renewal [21, 80,
81]. Retinoic acid is used to promote neural differentiation
through MAPK and integrin pathways [82].
Retinoic acid’s roles during embryogenesis has been
well documented. Retinoic acid promotes the expression
of genes involved in the development of central nervous
system, embryonal circulatory as well as heart asymmetry
[83]. Vitamin A-deficient embryos presented various con-
genital malformations, such as absence of eyes as well as
deficiencies in the central nervous system, skin, lungs and
heart [8486].
Retinoic acid also plays critical roles in cell fate deter-
mination in later stage of embryogenesis. For example, in
heart development, retinoic acid is involved in cardiac dif-
ferentiation. It modulates vascularization by suppressing the
gene expression of N-cadherin, Msx1 and TGFβ pathways;
It affects heart asymmetry through the inhibition of Nodal,
Snail and Pitx2 genes; It also promotes cell proliferation
and enhances BMP2 pathway by affecting the cardiogen-
esis transcription factor GATA4 [8794]. Based on retinoic
acid’s function in embryogenesis, it has been used to gener-
ate atrial cardiomyocytes [20]. In hematopoiesis, retinoic
acid enhances the exvivo maintenance and viability of
transplantable hematopoietic stem cells [95]. Retinoic acid
suppresses the proliferation of dormant hematopoietic stem
cells (HSCs), and prevents HSC differentiation to down-
stream cell types [96, 97]. As a result, retinoic acid helps
maintain the multipotency of HSCs, being enriched in these
cells compared to other multipotent progenitors [9799].
Furthermore, retinoic acid is also involved in germline dif-
ferentiation. Due to its interaction with BMP and NOTCH
pathways, retinoic acid’s targets are involved in four main
developmental stages of fetal germ cell development [82,
93, 100]. Retinoic acid increases the expression of germline
markers VASA, SCP3, TEKT1 and GDF9 [101], and pro-
motes the generation of tailed male gamete-like cells that
could generate offspring in mice [102].
Enzymes involved in retinoid acid production play
essential roles in embryogenesis. The oxidation of retinol
to retinal is the rate-limiting step in RA production, and the
enzymes RDH10 (short-chain dehydrogenase in charge of
the second oxidation of retinol) and DHRS3 (short-chain
dehydrogenase reductase in charge of reducing retinal to
retinol) are key in this process. Knockouts of these enzymes
result in developmental defects in craniofacial, heart and
limb patterning. RDH10-K.O. is lethal between E10.5 and
E14.5, and DHRS3-K.O. is lethal between E17.5 and E18.5
[103105]. Retinaldehyde dehydrogenase, which facilitates
the generation of retinoic acid from all-trans retinal, is a key
enzyme involved in cell fate determination [20, 66].
Although retinoic acid leads to ESC differentiation, it
is also paradoxically a potent promoter for somatic repro-
gramming. Somatic cells can be reprogrammed to induced
pluripotent stem cells (iPSCs) by the overexpression of tran-
scription factors, such as OCT4, KLF4, MYC and SOX2 [11,
106, 107]. The activation of retinoic acid pathway acceler-
ates reprogramming, while its removal suppresses repro-
gramming efficiency [108, 109]. The activation of retinoic
acid pathway is essential component in chemically induced
reprogramming without overexpressing transcription fac-
tors [110, 111]. Short-term treatment with retinoic acid is
reported to promote pluripotency of iPSCs by inhibiting
the canonical Wnt pathway, while positively modulating
AKT/mTOR signaling [112]. Additionally, retinol and RA
promote the transcription of Ten-eleven translocation (Tet)
proteins in naïve pluripotent stem cells, and the regulation
of Tet proteins by vitamin A is independent of vitamin C, a
known modulator of enzymatic activities (see more discus-
sions in “Vitamin C” section) [113]. In addition, retinoic
acid signaling is found to maintain the dormancy of HSCs
through cell cycle regulation [97].
Vitamin B3
Similar to vitamin A, vitamin B3 is also a family of com-
pounds including niacin (nicotinic acid), nicotinamide
(NAM) and nicotinamide riboside (NR). They are pre-
cursors of nicotinamide adenine dinucleotide (NAD) and
nicotinamide adenine dinucleotide phosphate (NADP) that
serve as cofactors or substrates in a wide range of metabolic
reactions [114, 115], so they are implicated in all metabolic
processes that utilize NAD or NADP (Fig.1). Because of
NAD’s importance in metabolism, there are both de novo
and salvage pathways for NAD synthesis from niacin, nicoti-
namide and NR (Fig.3). Nicotinamide is usually maintained
at around 100–200nM range in blood, while 16.6μM nico-
tinamide is supplied in DMEM/F12, which is sufficient to
sustain nutritional requirement of cells invitro (Table2).
C.Godoy-Parejo et al.
1 3
Nicotinamide has been utilized in clinical applications
(Table4). Nicotinamide ameliorates age-related macular
degeneration phenotypes [116]. It prevents hepatosteatosis
in obese mice while improving glucose metabolism and
increasing health span in mice [117]. These therapeutic
effects imply that nicotinamide could be involved in func-
tions beyond nutritional regulation.
Compared to regular culture for somatic cells, a higher
concentration of nicotinamide (5–10mM) are often used
in stem cell manipulations [19]. Nicotinamide in medium
can easily cross plasma membrane and translocate into
cytoplasm [19]. Nicotinamide was reported to promote cell
survival of hESCs. In differentiation, it promotes cardiomyo-
cyte differentiation, and facilitates the generation of endo-
crine pancreatic cells [118, 119]. Nicotinamide is also used
in the maintenance of somatic stem cells [120], as well as
organoid culture of different cell types [121123]. It is used
in the expansion of hematopoietic progenitors [124].
Nicotinamide is involved in various stem cell applica-
tions, but its exact molecular mechanism in each process is
De Novo Salvage
Neural Death
Oxidative Stress
Cardioprotection hPSC Survival
Kinase Inhibitor
Mesodermal specification/
Pancreatic progenitors
RPE differentiation
Hematopoietic progenitors
high concentration
physiological conditions
HSCs Survival
Proton Donor
Fig. 3 Vitamin B3 function in metabolism and signal transduction.
NAD+ is synthesized in humans by de novo and salvage pathways. In
the salvage pathway, the enzyme nicotinamide phosphoribosyltrans-
ferase (NAMPT) converts NAM (nicotinamide) to NMN (nicotina-
mide mononucleotide), which is then metabolized to NAD+. NAMPT
is the rate-limiting step in this process and is a crucial factor to main-
tain NAD+ levels. Nrk1 can directly phosphorylate NR (nicotinamide
riboside) to NMN, bypassing NAMPT, and NR can also be digested
into NAM. In the de novo pathway, QA (quinolinic acid) and niacin
are metabolized into NaMN (nicotinic acid mononucleotide) which
can be further catalyzed into NaAD (nicotinic acid adenine dinucleo-
tide). At high concentration, NAM functions as inhibitors in sirtuin,
PARP and kinase pathways
Table 4 Clinical applications of nicotinamide [269275]
Conditions Dose of nicotinamide
Acne 750mg/day
Vitamin B3 deficiency (Pellagra) 300–500mg/day
Diabetes 1.2g/mL/day
Hyperphosphatemia 0.5–1.75g/day
Larynx cancer 60mg/kg of
niacinamide/h before
inhaling carbogen
Skin cancer (other than melanoma) 0.5g niacinamide/day
Osteoarthritis 3g/day
Roles ofvitamins instem cells
1 3
still unclear. At high concentration, nicotinamide can inhibit
the activities of sirtuins, a family of protein deacetylases
that regulate epigenetic modification and potential cell fates
[125]. Nicotinamide is used to enrich CD34+ hematopoietic
progenitors as a SIRT1 specific inhibitor [124]. At the same
time, nicotinamide is also an inhibitor of poly(ADP-ribose)
polymerase (PARP) that is involved in cell death [126, 127].
It is thought to improve cell survival by inhibiting apoptosis.
Recently, nicotinamide was identified as a kinase inhibitor
at high concentration (millimolar range) [19]. Nicotinamide
targets multiple kinases that are involved in cell survival
and pluripotency. It binds and inhibits ROCK kinases, and it
suppresses cell death caused by ROCK activation after cell
individualization. Nicotinamide is also an inhibitor of casein
kinase 1 (CK1). The inhibition of CK1 leads to the exit of
self-renewal, and also promotes differentiation towards reti-
nal pigment epithelium [19]. It is foreseeable that nicotina-
mide could be involved in additional stem cell regulations as
a modulator in sirtuin, PARP and kinase pathways.
The concentration-dependent phenomena also exist in
some other nicotinamide derivatives, such as nicotinamide
mononucleotide (NMN) and NR. Recent studies show that
NMN and NR have functions beyond NAD synthesis. With
elevated concentration, NMN reverses vascular dysfunction
and oxidative stress, and promotes cardioprotection via gly-
colysis and acidic pH [128, 129]. NMN also protects against
cognitive impairment and neuronal death induced by the
inhibition of long-term potentiation (LTP) after Aβ1–42
oligomer treatment [130]. NR at elevated concentration
increases mitochondrial recycling and cell survival in hemat-
opoietic stem cells [131]. It also prevents aging, and extends
life span [132]. It is intriguing why nicotinamide derivatives
have such concentration-dependent effect. It would be inter-
esting to explore potential connections in these biological
Vitamin C
Vitamin C, or l-ascorbic acid (AA/LAA), is soluble in
water due to its sugar-like structure. Although ascorbic
acid is found at equal amounts in both isomeric states, l
and d-ascorbic acid, only LAA is chemically active. Ascor-
bic acid can be synthesized in plants and the majority of
animals (Fig.4, adapted from Linster’s and Schaftingen’s
review) [133]. In vertebrates, the last step of ascorbic acid
biosynthesis from glucose is the formation of 2-keto-gulono-
lactone which spontaneously enolizes into ascorbic acid. The
enzyme for this step, l-gulonolactone oxidase, is found inac-
tive in high primates, including humans, so human beings
have to take vitamin C from food sources [133, 134]. LAA
is not stable in nature due to its hydrogen ion, and acidic
pH will increase its stability. When exposed to light, it gets
oxidized to dehydroascorbic acid (DHA) [135]. In practice,
more stable LAA derivatives are used in cell culture, such
as magnesium ascorbyl phosphate (MAP) and ascorbyl 6
palmitate (AA6P) [136138].
Vitamin C is a potent antioxidant and reduces reactive
oxygen species (ROS), and it participates in various bio-
logical processes [139]. In addition, vitamin C acts as a
kinase inhibitor. When it is oxidized into dehydroascorbic
acid (DHA), it inhibits IκBα Kinase β and modulates NF-κB
signaling [140, 141]. Vitamin C also reduces ferric to fer-
rous iron, and increases its absorption in the intestine [142].
High doses of vitamin C can actually promote an oxi-
dative state in cancer cells, acting as a potential anti-can-
cer therapy [143145]. It is proposed that the anti-cancer
effect may be due to induction of ferroptosis, a form of pro-
grammed cell death related to vitamin E deficiency and lipid
peroxidation [146148]. High doses of ascorbic acid was
reported to regress Charcot-Marie-Tooth disease in mice, a
neuropathy with impairment in the myelination of periph-
eral nerves, due to the myelination effect of ascorbic acid
[149152]. The lack of Vitamin C is the trigger of a well-
known avitaminosis called scurvy, which if prolonged in
time can be fatal due to hemorrhages and impaired wound
healing [31].
Vitamin C plays critical roles in promoting PSC survival
and derivation. When hESCs are transitioned from mTeSR
medium to albumin-free and more defined condition, cells
die in the absence of vitamin C after a few days [50]. At the
same time, vitamin C also regulates the homeostasis of the
extracellular matrix [18]. It affects the folding and deposi-
tion of collagen proteins, which may have contributed to its
effect on hESC attachment and survival [27, 50, 153]. Dur-
ing reprogramming, ascorbic acid promotes reprogramming
in human and mouse cells [50, 154]. Vitamin C reduces cell
senescence during reprogramming by suppressing p53 [155,
156]. It was shown to act through a mechanism independ-
ent from its antioxidant role, and accelerates transcriptional
changes during reprogramming [154, 157]. Vitamin C also
influences cell survival in reprogramming through epige-
netic modulation. It is a cofactor for polyhydroxylates and
demethylases [158], and promotes demethylase activity on
shore CpG islands involved in tissue-specific DNA methyla-
tion and reprogramming [159, 160].
Besides its use for the maintenance of pluripotent stem
cells, vitamin C also impacts the differentiation of multiple
cell lineages. Vitamin C triggers mesoderm differentiation
of mouse embryonic stem cells [161]. It promoted myogen-
esis and osteogenesis, and inhibited adipogenesis. Vitamin
C inhibits neurogenesis to favor myogenesis through the
activation of the p38 MAPK/CREB pathway and chromatin
remodeling [161, 162]. It also promotes cardiac differen-
tiation and increases the proliferation of cardiac progenitor
cells by enhancing collagen synthesis [163].
C.Godoy-Parejo et al.
1 3
In addition to ESCs, vitamin C also regulates mesen-
chymal stem cell growth and differentiation [164166].
It suppresses hypoxia inducible factor 1 (HIF1α) activity
through two parallel pathways. Vitamin C suppresses HIFα
transcription, while activating HIF1α hydroxylase to break-
down HIF1α. Inhibition of HIF1α leads to mitochondrial
activation, affecting cell proliferation and metabolism [167].
MSCs cultured with vitamin C show upregulation of Oct4
and Sox2, without affecting the expression of MSC markers
such as CD105 and CD13 [168, 169]. Vitamin C in combi-
nation with TGFβ treatment was shown to promote MSC
differentiation toward vascular smooth muscle cell types
[170, 171]. Vitamin C also facilitates osteogenic differen-
tiation by increasing collagen secretion, since it is used as a
cofactor for enzymes that hydroxylate proline and lysine in
pro-collagen [171174]. Vitamin C also enhances chondro-
genic differentiation [175], and protects chondrocytes from
oxidative stress due to hydrogen peroxide (H2O2) [176].
Vitamin C is also beneficial to hematopoietic differen-
tiation and it has been used to promote the maturation of
T cells and NK cells from HSC-derived progenitors [177,
178]. Ascorbic acid is used to generate hematopoietic
Fig. 4 a Vitamin C regulation
in stem cells. Vitamin C, com-
monly referring to as l-ascorbic
acid, cannot be synthesized by
humans due to the lack of the
hydrolase gluconolactonase
enzyme. Similar to vitamin A,
it acts as an antioxidant and
tumor suppressor, and increases
the myelination of neurons. Its
effect on chromatin remodeling
and other epigenetics marks
allows it to affect reprogram-
ming of pluripotent stem cells,
but it is also necessary for the
culture of both embryonic and
mesenchymal stem cells. It pro-
motes hematopoiesis differen-
tiation and promotes mesoderm
lineages including cardiomyo-
cytes, bone and cartilage. The
effect of vitamin C on adipocyte
differentiation depends on
the platform and concentra-
tion. b Vitamin C regulation
of reprogramming. l-Ascorbic
acid facilitates the reaction
that converts ferric ions (Fe3+)
to ferrous ions (Fe2+). Fe2+ is
required for the activity of both
Tet proteins and JHDM histone
demethylases, and it is oxidated
into Fe3+ when α-KG and O2
are converted into succinate and
CO2 during DNA and histone
L-Ascorbic Acid
Stem Cell Survival/
hydrolase gluconolactonase
Inactive in high primates
Epigenetic Modulation/
L-Ascorbic Acid
DNA methylation
Deoxycytidine5mC 5hmC
O2+ α-KG
Succ + CO2
Fe2+ Fe3+
L-Ascorbic Acid
Lysine Lysine
O2+ α-KG
Succ + CO2
Fe2+ Fe3+
Histone methylation
Roles ofvitamins instem cells
1 3
stem cell progenitors (hemangioblasts) from hESCs [179].
Ascorbic acid concentration is high in human and mouse
hematopoietic stem cells (HSCs), and declines upon differ-
entiation. With the accumulation of intracellular ascorbic
acid, HSC frequency is limited, while leukemogenesis is
suppressed [180, 181].
Besides its antioxidant activity, vitamin C mainly acts
as an enzyme cofactor for the demethylation of DNA and
histone in stem cells (Fig.4b). Changes in DNA and histone
methylation are often associated with stem cell differentia-
tion and reprogramming [182184]. The methylation on the
fifth position of the pyrimidine ring of cytosine (5mC) is
the most common DNA modification, and its demethyla-
tion to 5-hydroxymethylcytosine (5hmC) is catalyzed by
Tet proteins [185187]. On the other hand, histone demeth-
ylation is carried out by histone demethylases such as the
Jumonji-C domain-containing family (JHDMs) [184, 188,
189]. Both Tet and JHDM proteins are vitamin C-depend-
ent, Fe2+/alpha-ketoglutarate-dependent hydroxylases
(Fe2+/α-KGDDs). During demethylation, Fe2+/α-KGDD
catalyzes the reaction that converts α-ketoglutarate (α-KG)
and O2 into succinate and CO2. Fe2+/α-KGDD activity
requires Fe2+ that is oxidized to Fe3+ in the process [148,
181, 190192]. Vitamin C reduces Fe3+ back to Fe2+ which
could then be utilized by Tet or JHDM in demethylation
again, while vitamin C itself is oxidized into dehydroascor-
bic acid (DHA) [113, 193]. Vitamin C influences the bio-
logical outcome of Tet-mediated DNA demethylation, and
promotes the demethylation of histones such as H3, H3K9,
H3K36 and H3K27 [194]. Collectively, vitamin C enhances
the efficiency of somatic programming [154]. In addition,
vitamin C also impacts stem cell differentiation. Vitamin
C improves HSC differentiation by modulating Tet activity
[180, 181], and it also increases the expression of key genes
in dopaminergic neurons in the fetal brain [195], as well as
trophectoderm genes like Cdx2, Eomes and Elf2 in the dif-
ferentiation of mouse embryonic stem cells [196].
Vitamin E
Since the discovery of α-tocopherol in 1922 [197], vitamin
E has been extensively studied and become one of the most
commonly consumed vitamins. There are eight known natu-
ral isoforms of vitamin E, including four tocopherols and
four tocotrienols, each designated as α, β, γ and δ based
on the position of methyl groups on the chromanol ring
[198200]. Vitamin E exists in almost all the tissues in
the human body, with highest levels in the adipose tissue
and adrenal gland [200]. Early studies on vitamin E mostly
focused on α-tocopherol, the most abundant vitamin E iso-
form [200]. Compared to the other isoforms, α-tocopherol
has higher bioavailability and longer retention time, due
to its preferential incorporation into lipoproteins by alpha-
tocopherol transfer protein (α-TTP) in the liver [199, 201].
It is also the isoform commonly provided in dietary sup-
plements [199]. In recent years, non-α-tocopherols have
received increasing attention, and the tocotrienols are
reported to be superior over tocopherols in many clinical
applications [201203]. Synthetic forms of vitamin E and its
chemically modified analogs, such as trolox [204], tocoflexol
[205] and esters of vitamin E [206209] are also widely used
for improved bioavailability and stability.
Vitamin E is a lipid soluble, chain-breaking antioxidant,
capable of neutralizing free radicals and terminating chain
reactions in the oxidation of polyunsaturated fatty acids. It
is one of the major antioxidants in the human plasma [210].
Due to its lipid solubility, vitamin E effectively protects
against oxidative damage from lipid peroxidation in the
membrane as well as in lipid vesicles, but is less effective
against damage from aqueous free peroxyl radicals [210,
In addition to its antioxidant role, vitamin E also mod-
ulates cellular signal transduction through kinases, phos-
phatases, lipid mediators and transcription factors [35, 212].
α-Tocopherol inhibits protein kinase C (PKC), while other
vitamin E isoforms were reported to have no influence or
opposing effect [213215]. Regulation of PKC by vitamin
E leads to changes in cell proliferation, adhesion, gene
expression and downstream signal transduction [35, 213,
216, 217]. Another important target of vitamin E is protein
kinase B (PKB/AKT), which plays a key role in cell sur-
vival. Vitamin E may activate or inhibit PI3K/AKT pathway
and cell survival in a cell type-specific manner [218221].
Other signaling pathways regulated by vitamin E include
ERK [219], p38 MAPK [222] and Wnt signaling [223]. Due
to its influence on membrane composition, vitamin E can not
only directly or indirectly activate/inhibit its targets, but also
change specific structural features of the plasma membrane
(such as lipid rafts), which may be involved in the membrane
translocation or activation of signaling molecules [212].
Vitamin E was frequently used in primary cell culture to
prevent cell death and preserve cell function after exposure
to stress conditions, and both antioxidant and signal trans-
duction modulating mechanisms may be involved. For exam-
ple, vitamin E treatment during enzymatic dissociation pro-
tected rat mammary epithelial cells against oxidative damage
and improved survival [224]. γ-Tocotrienol was reported to
enhance AKT phosphorylation in intestinal tissue following
total body irradiation, thereby protecting the tissue against
damage by radiation [221]. Low micromolar concentrations
of α-tocopherol suppressed the rise of metalloproteinase 1
(MMP-1) expression in UVA-irradiated fibroblasts, sug-
gesting a photoprotective effect [225]. In an endothelial cell
model for type I diabetes, 20mg/L α-tocopherol showed
protective effects against endothelial dysfunction caused by
C.Godoy-Parejo et al.
1 3
hyperglycemia [226]. In some studies, high concentrations
(200–2500µmol/L) of vitamin E were used for cell culture
[227229], far exceeding the reported plasma vitamin E lev-
els ranging from 15 to 27µmol/L [230233].
Commercial cell culture supplements containing vitamin
E are available. The B-27 supplement is widely used for neu-
ronal cell culture [234, 235], and chemically defined lipid
concentrate is used to support mammalian and insect cell
culture in place of fetal bovine serum [236]. The isoform of
vitamin E supplied in these supplements are α-tocopherol
or α-tocopherol acetate in low micromolar concentrations.
As a potent antioxidant, vitamin E was reported to be pro-
tective for stem cells and progenitor cells which are sensitive
to oxidative stress. Treatment with α-tocopherol protected
mesenchymal stem cells (MSCs) against H2O2-induced
apoptosis and promoted MSC survival via the AKT path-
way [220, 237]. Similarly, trolox was reported to enhance the
proliferation of human dental pulp stem cells under oxygen
tension [238]. α-tocopherol also promoted the survival of
cultured human neural progenitors, and the effect was abol-
ished by inhibitors of PI3K/AKT and Src signaling [239].
This is consistent with invivo studies using mouse models,
in which vitamin E deficiency or impairment of its uptake
resulted in neural tube defects [240, 241].
In addition to affecting cell survival, vitamin E was also
reported to affect differentiation of stem cells as a free radi-
cal scavenger. Reactive oxygen species (ROS) were pro-
posed to participate in cellular signaling and regulate embry-
onic stem cell (ESC) differentiation, and vitamin E typically
antagonizes the ROS effects. Arachidonic acid, the precursor
of prostaglandins and leukotrienes, was reported to promote
the generation of vascular progenitor cells from mouse ESC
embryoid bodies. ROS was elevated in the process, and
trolox treatment from day 3 to day 10 abolished the effect of
arachidonic acid on differentiation [242]. In another study,
electrical field treatment stimulated endothelial differentia-
tion of mouse ESCs through a mechanism involving ROS,
and trolox treatment inhibited its effect [243]. In cardiac
differentiation from mouse ESCs, treatment with valproic
acid from day 3 to 7 was reported to inhibit embryoid body
growth and suppress cardiomyocyte differentiation while
increasing ROS. Co-administration of trolox antagonized the
inhibitory effect and restored cardiomyocyte differentiation
[244]. In contrast, icariin treatment from day 5 to 16 of car-
diac differentiation, which elevated ROS and induced ERK/
p38 phosphorylation, significantly enhanced cardiac differ-
entiation, and vitamin E treatment decreased the promoting
effect by half [245]. Similarly, elevated intracellular ROS
by cardiotrophin-1 (CT-1, from day 7 on) is associated with
improved cardiomyocyte differentiation and increased Ki-67
expression, suggesting better cardiomyocyte proliferation.
Vitamin E abolished these effects as well through a mecha-
nism involving Jak/Stat and ERK pathways [246]. Taken
together, vitamin E can play regulatory roles during ESC
differentiation toward multiple lineages, potentially through
a mechanism involving ROS generation and activation of
relevant signaling pathways. The exact effect may depend
on the setting of differentiation and the timing of treatment.
The functions of vitamin E are summarized in Fig.5.
Coordinated vitamin actions instem cell
Each vitamin has its distinctive role in biochemical pro-
cesses, and many of them work together to carry out critical
cellular functions. For example, the generation of acetyl-
CoA requires B1, B2, B3 and B5, which is essential for both
somatic and stem cells. Many other biological processes also
demand collaborative actions of multiple vitamins, and some
of them are especially important to stem cells.
Epigenetic regulation is essential for self-renewal and cell
fate determination [247]. DNA and histone methylation are a
key modification, and it is responsive to nutrition and meta-
bolic changes. Appropriate epigenetic regulation is essential
for pregnancy and embryonic development. Vitamin B12, B9,
and B6 are key coenzymes in one carbon metabolism and
can synergistically influence DNA and histone methylation
[248, 249]. One carbon metabolism involves the donation
of carbon units from amino acids for utilization in various
biochemical reaction. In the folate (B9) cycle, a carbon unit
AntioxidantSignal transduction modulator
Cell Survival
Radiation Damage
Stem Cell Differentiation
Fig. 5 Vitamin E in stem cell culture. Vitamin E is a potent lipid-
soluble antioxidant, and is capable of protecting stem cells and pro-
genitor cells against oxidative damage. In addition, vitamin E can
modulate signaling pathways, including the PI3K/AKT pathway, to
promote survival and proliferation of cells in culture. The impact of
vitamin E on ESC differentiation is mainly mediated through ROS
levels. Whether vitamin E can directly modulate signal transduction
events involved in cell fate determination has not been reported
Roles ofvitamins instem cells
1 3
produced from the conversion of serine to glycine is trans-
ferred to tetrahydrofolate (THF) by serine hydroxymethyl-
transferase (SHMT), a vitamin B6-dependent enzyme. The
resulting 5,10-methylene-THF is important for nucleotide
synthesis. In the methionine cycle, vitamin B12 serves as a
coenzyme in the conversion of homocysteine to methionine
by accepting a carbon unit from the folate cycle. Methionine
is further converted to S-adenosylmethionine (SAM) [250,
251], which is the main methyl group donor for the methyla-
tion of proteins, DNA, RNA and lipids [185].
The combined actions of vitamins are also reflected in
multiple stem cell media containing vitamin combinations
(Table2), and are utilized in some stem cell protocols [252].
Concluding remarks
Vitamins are deeply involved in various basic metabolic
and signaling processes, and many of them are required for
normal functions in specific stem cells. Besides the con-
ventional approach of stem cell modulation through growth
factor signaling pathways, vitamin modulation could become
a critical approach to improve stem cell maintenance and
downstream differentiation. Studies on vitamins such as A,
B3 and C have shown that vitamin-dependent pathways are
effective targets in stem cell manipulation. However, most
vitamins have not been systematically explored in different
stem cell studies. Considering that specific cell types rely
on distinctive combinations of vitamins, it is possible that
more stem cell applications could be developed using differ-
ent vitamin formulations in media. At the same time, stem
cell culture also provides a unique platform to study vita-
min function in human embryogenesis. Following the recent
discoveries of vitamin-related molecular mechanisms, more
novel mechanisms could be identified in stem cell models.
We believe that vitamin study in stem cell research will
lead to new modulations to improve stem cell applications,
and help realize their great potentials in basic research and
regenerative medicine.
Acknowledgements This work was supported by the Science and
Technology Development Fund of Macau SAR (FDCT/131/2014/
A3, FDCT/056/2015/A2 and FDCT/0059/2019/A1) and University of
Macau Multiyear Research Grant (MYRG2018-00135-FHS).
Compliance with ethical standards
Conflict of interest The authors declare no conflict of interest.
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... The vitamins are chemical compounds that cannot be synthesized by the host or can only in minimum amounts and are essential for a wide variety of metabolic pathways. The word vitamin derives from Latin "vita" meaning "life," which denotes its essential roles for good health and well-being of humans [63]. They have a great diversity of chemical functional groups. ...
... For the past 60 years in numerous studies in cell cultures and laboratory animals have provided evidence indicating that the vitamins can modulate gene expression, proliferation and differentiation cell, and signal transduction at supraphysiological concentrations [63,121,134]. Additionally, vitamins at pharmacological concentrations demonstrated their benefits in MetS-related diseases. ...
... Another explanation is related with the polymorphisms influence vitamin binding protein levels and absorption, because there could be a considerable ethnic variation in their allele frequencies and affect the levels of bioavailable vitamin [8,109,166]. It is also important to consider that many vitamins (B1, B2, B5, B6, B7, B9, B12, and K vitamins) are synthesized by gut bacteria [63] and it has been shown that the microbiota is altered in individuals with diseases related to metabolic syndrome [53]. Therefore, the interaction of vitamins with microbiota could also be considered since vitamins with supraphysiological concentrations could modify the type and amount by regulating genes and signal transduction pathways in them as they do in mammals and plants. ...
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The prevalence of metabolic syndrome and cardiovascular disease has increased and continues to be the leading cause of mortality worldwide. The etiology of these diseases includes a complex phenotype derived from interactions between genetic, environmental, and nutritional factors. In this regard, it is common to observe vitamin deficiencies in the general population and even more in patients with cardiometabolic diseases due to different factors. Vitamins are essential micronutrients for cellular metabolism and their deficiencies result in diseases. In addition to its role in nutritional functions, increasingly, vitamins are being recognized as modulators of genetics expression and signals transduction, when consumed at pharmacological concentrations. Numerous randomized preclinical and clinical trials have evaluated the use of vitamin supplementation in the prevention and treatment of metabolic syndrome and cardiovascular disease. However, it is controversy regarding its efficacy in the treatment and prevention of these diseases. In this review, we investigated chemical basics, physiological effect and recommended daily intake, problems with deficiency and overdose, preclinical and clinical studies, and mechanisms of action of vitamin supplementation in the treatment and prevention of metabolic syndrome and cardiovascular disease.
... Anti-inflammatory and antioxidant, improving membrane fluidity Calder, 2016;Burchakov et al., 2017;Godhamgaonkar et al., 2020;Sundaram et al., 2020 Folate (Vitamin B9) Antioxidant activity and plays a role in mTOR signaling Williams et al., 2011;Rosario et al., 2017 Vitamin E Antioxidant activity and modulating protein-membrane interaction Howard et al., 2011;Zingg, 2015 Choline Generates phosphatidylcholine and prevents intramembranous material leakage Penry and Manore, 2008;Godoy-Parejo et al., 2020; Zinc Binding site of copper and iron, membrane rheology, cell signaling, enzyme activities, lipid composition maintenance Verstraeten et al., 2004;Wilson et al., 2017 apoptosis could be activated, which, if it progresses, may lead to necroptosis, resulting in abnormal placentation and vascular remodeling and therefore preeclampsia (Raguema et al., 2020;Zhang et al., 2020). Folic acid is crucial to healthy pregnancy as it contributes to angiogenesis and vasculogenesis via a nitric oxide-dependent mechanism, DNA methylation, antioxidant protection, and endothelial-dependent vascular relaxation. ...
... Deficiency of choline may cause intramembranous material to leak into extracellular fluid as the cell integrity is compromised. It is also involved in synthesis of lipoprotein, acetylcholine, and homocysteine (Penry and Manore, 2008;Godoy-Parejo et al., 2020). Moreover, an in vivo study showed that reduction of folate is associated with the increased breakdown of phosphatidylcholine and glycerophosphocholine (GPC), due to a high demand for the methyl group (Chew et al., 2011). ...
... Vitamin B2 is essential for complexes I and II building blocks and is involved in the TCA cycle during fatty acid oxidation. Vitamin B3 is the precursor (Wesselink et al., 2019;Godoy-Parejo et al., 2020). Low levels of folic acid is associated with oxidative damage, altered antioxidant enzymatic activities, decrease in mtDNA biogenesis, and mitochondrial oxidative deterioration. ...
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Preeclampsia is one of the most common obstetrical complications worldwide. The pathomechanism of this disease begins with abnormal placentation in early pregnancy, which is associated with inappropriate decidualization, vasculogenesis, angiogenesis, and spiral artery remodeling, leading to endothelial dysfunction. In these processes, appropriate cellular deaths have been proposed to play a pivotal role, including apoptosis and autophagy. The proper functioning of these physiological cell deaths for placentation depends on the wellbeing of the trophoblasts, affected by the structural and functional integrity of each cellular component including the cell membrane, mitochondria, endoplasmic reticulum, genetics, and epigenetics. This cellular wellness, which includes optimal cellular integrity and function, is heavily influenced by nutritional adequacy. In contrast, nutritional deficiencies may result in the alteration of plasma membrane, mitochondrial dysfunction, endoplasmic reticulum stress, and changes in gene expression, DNA methylation, and miRNA expression, as well as weakened defense against environmental contaminants, hence inducing a series of inappropriate cellular deaths such as abnormal apoptosis and necrosis, and autophagy dysfunction and resulting in abnormal trophoblast invasion. Despite their inherent connection, the currently available studies examined the functions of each organelle, the cellular death mechanisms and the nutrition involved, both physiologically in the placenta and in preeclampsia, separately. Therefore, this review aims to comprehensively discuss the relationship between each organelle in maintaining the physiological cell death mechanisms and the nutrition involved, and the interconnection between the disruptions in the cellular organelles and inappropriate cell death mechanisms, resulting in poor trophoblast invasion and differentiation, as seen in preeclampsia.
... The role of vitamin A in embryonic stem cell reprograming has been extensively studied, and it is mediated by TET demethylases, which remove DNA methylation marks and induce DNA demethylation. Retinoic acid is capable of inhibiting DNMTs, thereby, resulting in the transcriptional suppression or hypomethylation of DNA [37]. Histone alteration is another epigenetic procedure affected by RA. ...
Air pollution has emerged as a serious threat to human health due to close association with spectrum of chronic ailments including cardiovascular disorders, respiratory diseases, nervous system dysfunctions, diabetes and cancer. Exposure to air-borne pollutants along with poor eating behaviours and inferior dietary quality irreversibly impacts epigenomic landscape, leading to aberrant transcriptional control of gene expression which is central to patho-physiology of non-communicable diseases. It is assumed that nutriepigenomic interventions such as vitamins can control such adverse effects through their immediate action on mitochondrial epigenomic-axis. Importantly, the exhaustive clinical utility of vitamins-interceded epigenetic synchronization is not well characterized. Therefore, improving the current limitations linked to stability and bioavailability issues in vitamin formulations is highly warranted. The present review not only sums up the available data on the role of vitamins as potential epigenetic modifiers but also discusses the importance of nano-engineered vitamins as potential epidrugs for dietary and pharmacological intervention to mitigate the long-term effects of air pollution toxicity.
... Vitamins B1 and B3 are involved in glycolysis, while B1, B2, B3, B5 and B7 are important for the TCA cycle and oxidative phosphorylation. Lipid synthesis and metabolism require the actions of B2, B3, B5 and B7; Vitamins B3, B6, B9 and B12 are involved in amino acid metabolism [37]. All of them are usually supplied in the base medium for hPSCs. ...
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All living organisms need energy to carry out their essential functions. The importance of energy metabolism is increasingly recognized in human pluripotent stem cells. Energy production is not only essential for cell survival and proliferation, but also critical for pluripotency and cell fate determination. Thus, energy metabolism is an important target in cellular regulation and stem cell applications. In this review, we will discuss key factors that influence energy metabolism and their association with stem cell functions.
... Although the necessity of very low concentrations of this component, the presence of VITs in vitro is key and their absence may lead to decrease cell growth, death, or loss of function. Therefore, we add vitamins to our BACTs in order to improve local cell homeostasis and survival [47]. ...
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Regenerative medicine and surgery is a rapidly expanding branch of translational research in tissue engineering, cellular and molecular biology. To date, the methods to improve cell intake, survival, and isolation need to comply with a complex and still unclear regulatory frame, becoming everyday more restrictive and often limiting the effectiveness and outcome of the therapeutic choices. Thus, the authors developed a novel 360° regenerative strategy based on the synergic action of several new components called the bioactive composite therapies (BACTs) to improve grafted cells intake, and survival in total compliance with the legal and ethical limits of the current regulatory frame. The rationale at the origin of this new technology is based on the evidence that cells need supportive substrate to survive in vitro and this observation, applying the concept of translational medicine, is true also in vivo. Bioactive composite mixtures (BACMs) are tailor-made bioactive mixtures containing several bioactive components that support cells’ survival and induce a regenerative response in vivo by stimulating the recipient site to act as an in situ real bioreactor. Many different tissues have been used in the past for the isolation of cells, molecules, and growth factors, but the adipose tissue and its stromal vascular fraction (SVF) remains the most valuable, abundant, safe, and reliable source of regenerative components and particularly of adipose-derived stems cells (ADSCs). The role of plastic surgeons as the historical experts in all the most advanced techniques for harvesting, manipulating, and grafting adipose tissue is fundamental in this constant process of expansion of regenerative procedures. In this article, we analyze the main causes of cell death and the strategies for preventing it, and we present all the technical steps for preparing the main components of BACMs and the different mixing modalities to obtain the most efficient regenerative action on different clinical and pathological conditions. The second section of this work is dedicated to the logical and sequential evolution from simple bioactive composite grafts (BACGs) that distinguished our initial approach to regenerative medicine, to BACTs where many other fundamental technical steps are analyzed and integrated for supporting and enhancing the most efficient regenerative activity. Level of Evidence: Not gradable
... It is present in cell membranes and plays a significant preventive role in the oxidative damage of molecules such as DNA or lipids [20,68]. α-Tocopherol neutralizes free radicals and breaks the chain reaction in the oxidation of the polyunsaturated fatty acids [69]. This activity is associated with the non-esterified hydroxyl groups of the chromanol ring. ...
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Tocopherols and tocotrienols are natural compounds of plant origin, available in the nature. They are supplied in various amounts in a diet, mainly from vegetable oils, some oilseeds, and nuts. The main forms in the diet are α- and γ-tocopherol, due to the highest content in food products. Nevertheless, α-tocopherol is the main form of vitamin E with the highest tissue concentration. The α- forms of both tocopherols and tocotrienols are considered as the most metabolically active. Currently, research results indicate also a greater antioxidant potential of tocotrienols than tocopherols. Moreover, the biological role of vitamin E metabolites have received increasing interest. The aim of this review is to update the knowledge of tocopherol and tocotrienol bioactivity, with a particular focus on their bioavailability, distribution, and metabolism determinants in humans. Almost one hundred years after the start of research on α-tocopherol, its biological properties are still under investigation. For several decades, researchers’ interest in the biological importance of other forms of vitamin E has also been growing. Some of the functions, for instance the antioxidant functions of α- and γ-tocopherols, have been confirmed in humans, while others, such as the relationship with metabolic disorders, are still under investigation. Some studies, which analyzed the biological role and mechanisms of tocopherols and tocotrienols over the past few years described new and even unexpected cellular and molecular properties that will be the subject of future research.
Metabolism has been studied mainly in cultured cells or at the level of whole tissues or whole organisms in vivo. Consequently, our understanding of metabolic heterogeneity among cells within tissues is limited, particularly when it comes to rare cells with biologically distinct properties, such as stem cells. Stem cell function, tissue regeneration and cancer suppression are all metabolically regulated, although it is not yet clear whether there are metabolic mechanisms unique to stem cells that regulate their activity and function. Recent work has, however, provided evidence that stem cells do have a metabolic signature that is distinct from that of restricted progenitors and that metabolic changes influence tissue homeostasis and regeneration. Stem cell maintenance throughout life in many tissues depends upon minimizing anabolic pathway activation and cell division. Consequently, stem cell activation by tissue injury is associated with changes in mitochondrial function, lysosome activity and lipid metabolism, potentially at the cost of eroding self-renewal potential. Stem cell metabolism is also regulated by the environment: stem cells metabolically interact with other cells in their niches and are able to sense and adapt to dietary changes. The accelerating understanding of stem cell metabolism is revealing new aspects of tissue homeostasis with the potential to promote tissue regeneration and cancer suppression. The metabolism of somatic stem cells must be regulated to meet their specific needs, to enable long-term maintenance as well as their activation, proliferation and subsequent differentiation. Better understanding of metabolic regulation in stem cells will open new opportunities to manipulate stem cell function, with potential applications in tissue regeneration and cancer prevention.
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Regenerative Medicine and Surgery is a rapidly expanding branch of translational research in tissue engineering, cellular and molecular biology. To date, the methods to improve cell intake, survival and isolation need to comply with a complex and still unclear regulatory frame, becoming everyday more restrictive and often limiting effectiveness and outcome of the therapeutic choices. Thus, the author developed a novel regenerative strategy, based on the synergic action of several bio-active components, called the Bio-Active Composite Therapies (BACTs) to improve grafted cells intake and survival in total compliance with the legal and ethical limits of the current regulatory frame. The rationale at the origin of this new technology is based on the evidence that cells need supportive substrate to survive in vitro and this observation, applying the concept of translational medicine, is true also in vivo. Many different sources have been used in the past for MSCs, molecules and growth factors (GF) isolation and extraction, but the Adipose Tissue and its Stromal Vascular Fraction (SVF) definitely remains the most valuable, abundant, safe and reliable. Bio-Active Composite Mixtures (BACMs) are tailor-made injectable “cocktails” containing several bio-active components to support cells survival and induce a strong regenerative response in vivo by stimulating the recipient site to act as an in-situ real Bioreactor. In this article, the author analyze the main causes of cell’s death and the strategies for preventing it, and outline all the technical steps for preparing the main components of BACMs and the different mixing modalities to obtain the most efficient regenerative action on different clinical and pathological conditions in several surgical specialties. Orthopedic Surgery is definitely the one that most can benefit of these new therapeutic strategies. The final part of this work is anticipating the logical and sequential evolution toward other fundamental technical steps for further supporting and enhancing the most efficient regenerative activity.
A balanced diet with adequate nutrient intake is vital for normal fetal kidney development to occur. In contrast, maternal malnourishment during pregnancy impairs fetal kidney development due to a deficiency in nutrients that the mother and growing fetus have access to. Nutritional deficiency not only reduces the structural building blocks needed for nephron generation but also impacts various signaling pathways, growth factors, epigenetic modifications, gene expression, and oxidative stress that collectively blunts nephrogenesis. These effects of nutritional deficiency consequently reduce the stem cell pool that is available for nephron building by altering their self-renewal, survival, and patterned differentiation. As a result, the fetus is born following maternal malnourishment with significantly fewer nephrons, underdeveloped kidneys, potential kidney dysfunction, and a lifelong increased susceptibility to kidney and cardiovascular disease. Discussed in this chapter are the specific affects that vitamins, minerals, amino acids, lipids, and fatty acids have on stem cells, nephrogenesis, and kidney development.
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The escalating prevalence of coronavirus disease 2019 (COVID-19) worldwide, with an increased rate of morbidity and mortality, highlights an urgent need to develop more effective therapeutic interventions. Despite the authorized treatment against COVID-19 by the European Union (EU), the safety and effectiveness of this therapeutic strategy for a wide variety of patients have remained a significant challenge. In this respect, micronutrients such as vitamins and minerals, as essential factors, can be considered for improving the function of the immune system and accelerating the treatment procedure. Dietary supplements can attenuate vascular and inflammatory manifestations related to infectious diseases in large part due to their anti-inflammatory and antioxidant properties. Recently, it has been revealed that poor nutritional status may be one of the notable risk factors in severe COVID-19 infections. In the current review, we focus on the micronutrient therapy of COVID-19 patients and provide a comprehensive insight into the essential vitamins/minerals and their role in controlling the severity of the COVID-19 infection. We also discuss the recent advancements, challenges, negative and positive outcomes in relevance to this approach.
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The erasure of epigenetic modifications across the genome of somatic cells is an essential requirement during their reprogramming into induced pluripotent stem cells (iPSCs). Vitamin C plays a pivotal role in remodeling the epigenome by enhancing the activity of Jumonji-C domain-containing histone demethylases (JHDMs) and the ten-eleven translocation (TET) proteins. By maintaining differentiation plasticity in culture, vitamin C also improves the quality of tissue specific stem cells derived from iPSCs that are highly sought after for use in regenerative medicine. The ability of vitamin C to potentiate the activity of histone and DNA demethylating enzymes also has clinical application in the treatment of cancer. Vitamin C deficiency has been widely reported in cancer patients and has recently been shown to accelerate cancer progression in disease models. Therapies involving high-dose vitamin C administration are currently gaining traction in the treatment of epigenetic dysregulation, by targeting aberrant histone and DNA methylation patterns associated with cancer progression.
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The media formulations necessary for deriving and sustaining organoids from epithelial tissues such as prostate, colon, gastric, liver, pancreas, and others have been established. Critical components of organoid media are a set of growth factors that include R-spondins and BMP signalling antagonists such as Noggin or Gremlin 1. Currently, the practical limitations for formulating organoid media of reproducible potency and larger-scale media production that have hampered further technological applications of organoid technology include: the cost of growth factors such as R-spondins and Gremlin 1/Noggin and their production as defined specific activities free of contaminants that may affect organoid growth. Here we report the production of highly pure recombinant Gremlin 1 and R-spondin 1 from bacterial expression for use in organoid media. We detail the workflow for Gremlin 1 and R-spondin 1 expression, purification, quantification of cellular activity, quality control and use in media formulated for culturing organoids derived from a number of tissues. The development of precisely formulated, cost-effective media of defined specific activity will engender the development of novel applications for organoid technology.