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Abstract

Umbilical cord blood (UBC) can be viewed as the most promising source of stem cells, in which collection cost is minimal and its benefits are immense. The cord blood is used to treat malignant and nonmalignant diseases; this is due to its progenitor characteristics know as stem cells.Its properties of being, immunologically immature and high plasticity has made it superior to other sources of stem cells. The stem cells collected from cord blood have neutral differentiation capabilities which allow medical professionals to produce functional neural cells from these stem cells.Cord Blood Banking (CBB) is the storing of the umbilical cord blood which is collected immediately after the delivery of the baby. Great care and concern are needed for proper storage of these progenitor cells, hence cord blood banks come into the play, they are of 3 types which are: public, private and direct donation banks.Clinical trials are still at its very early stages having abundances to still be uncovered but results were obtained have demonstrated high potential and more scope towards effective development therapies and treatments for rare disorders.
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Umbilical Cord Blood Banking and its Therapeutic Uses
Nivethika Sivakumaran
1
, Imesha Rashmini Rathnayaka
1
, Rashida Shabbir
1
, Sasini Sandareka Wimalsinghe
1
,
J. A. Sumalimina Jayakody
1
, Mahisha Chandrasekaran
1
1
International College of Business and Technology, Biomedical Science Department, No 502A, R. A. De Mel
Mawatha, Sri Lanka
*Corresponding author
Nivethika Sivakumaran
Email: snivethika25@gmail.com
Fax: +94 11 4203170
Author Nivethika Sivakumaran E-mail: snivethika25@gmail.com,Mobile No. - +94771048206
Author Imesha Rashmini Rathnayaka E-mail: imasharashmini@gmail.com, Mobile No. -+94775349616
Author Rashida Shabbir E-mail: rashida7755@gmail.com, mobile No. - +94774319976
Author SasiniSandarekaWimalsingheE-mail: wsasini@yahoo.com,Mobile No. - +94764359634
Author J. A. SumaliminaJayakodyE-mail: sumalimina19@gmail.com, Mobile No. - +94768925901
Author MahishaChandrasekaranE-mail: mahishachandrasekaran@gmail.com, Mobile No. - +94752970463
*Funding Organization: This Literature Review article is funded by International College of Business and
Technology,No 502A, R. A. De Mel Mawatha, Bambalapitiya, Colombo 04, Sri Lanka.
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Abstract
Umbilical cord blood (UBC) can be viewed as the most promising source of stem cells, in which collection
cost is minimal and its benefits are immense. The cord blood is used to treat malignant and nonmalignant
diseases; this is due to its progenitor characteristics know as stem cells.
Its properties of being, immunologically immature and high plasticity has made it superior to other sources of
stem cells. The stem cells collected from cord blood have neutral differentiation capabilities which allow
medical professionals to produce functional neural cells from these stem cells.
Cord Blood Banking (CBB) is the storing of the umbilical cord blood which is collected immediately after the
delivery of the baby. Great care and concern is needed for proper storage of these progenitor cells, hence
cord blood banks come into the play, they are of 3 types which are: public, private and direct donation banks.
Clinical trials are still at its very early stages having abundances to still be uncovered but results being
obtained have demonstrated high potential and more scope towards effective development therapies and
treatments for rare disorders.
Key words: umbilical cord blood, germ line layer, progenitor characteristics, cord blood banking, stem cells,
pluripotent, therapeutic uses
Abbreviations: UCB-umbilical cord blood; CBB- cord blood banking; EPCs- endothelial progenitor cells;
MSC- mesenchymal stromal cells; USSC- unrestricted stomatic stem cells; VSEL- very small embryonic-like
stem cells; MLPC- multi lineage progenitor cells; DNA- deoxyribose nucleic acid; HES- hydroxgethyl starch;
RCB- residual cancer burden; HLA - human leukocyte antigen; NC- natural killers; WBC- white blood cells;
NPBI- non protein bound iron; NC- natural cell; GvHD- graft vs host disease; MNCS- mono nucleated cells;
RBC- red blood cells; TNC- total nucleated cell; CD133,CD34,CD45- commonly used markers of
Hematopoetic progenitor cells and endothelial cell (Cluster of Differentitation; GMO- granulocyte
macrophage.
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1. Introduction
The umbilical cord is thin, long tube-like structure highly composed of muscle that is needed to form a
connection between the fetus and the placenta, in the mother’s uterus. To provide sufficient and effective
circulation of the blood to the growing fetus, the umbilical cord has one vein which carries oxygenated rich
blood and two arteries which carry deoxygenated blood. These 3 blood vessels coil around the vein in a helical
configuration to form the umbilical cord (Sadler, 2006)
After birth, the blood in the umbilical cord can be collected in two ways, either by a syringe or bag method
(Phuc Van Pham, 2014). The cord blood is used as an alternative to bone marrow to restore immunological
dysfunctions and for transplantations. This blood is found and collected from the umbilical cord in new born
infants (Weiss, 2006).
Present statistics have proved that nearly 80 diseases can be completely cured using umbilical cord blood stem
cells and over 50 000 transplants have been successfully carried out worldwide. Some of the diseases cured
are cancers and blood disorders which had so far always resulted fatal to many affected (Roura, 2015).
The properties of stem cells remain unknown, but their importance is beyond description. The cells in the
umbilical cord are unaffected by the external environment, and can effectively differentiate into many types of
cells. Life threatening disorder such as thalassemia can be cured with this stem cell, around the world there are
27 million babies affected with blood disorders which can now be cured with the use of the umbilical cord
stem cells. The umbilical cord blood is used to cure disorders and diseases for an individual to whom the
umbilical cord belongs to, therefore it is assured that there is no mismatching or rejection by the body, like in
many cases, people with rare blood groups find it difficult to find a matching donor. This major problem is
solved when umbilical cord blood stem cells are used. There is no counter effect after the treatment. Another
important factor to consider is that, if harmful diseases are not cured over generations, the threat is higher and
become public concern. Resulting in the wide spreading of the diseases and increase in numbers affected.
When such a disease is cured with umbilical cord blood stem cells, the effect of it being passed on is reduced
and therefore works towards making the disease endemic (Carroll, 2015).
2. Extraction and preservation of umbilical cord blood
There are different methods for both preservation and extraction, but parents are not often given the choice of
choosing what they like. Whether it be donating for a government hospital or storing in a private cord blood
for personal medicinal purposes.
The sole purpose of processing is to separate stem cells from the cord blood so that a sample is produced that
can be used safely (Cord Blood Banking How Baby’s Cells are Extracted & Stored? | LifeCell, 2017).The
separation and the processing of UCB samples that are present in a greater volume intended for storage in cord
blood banks use a partially automated system to make sure that the large numbers of samples are processed
efficiently. A closed system is usually used to reduce the risk of any bacterial infection due to a contamination.
This processing method allows the recovery of nucleated cells and progenitors adequately to enable the
engraftment procedure(Saunders Comprehensive Veterinary Dictionary, 2016).
Early studies have shown that the use of density gradient techniques to separate cord blood leads to the loss of
mononuclear cells. This proves that cord blood should be stored in an unseparated manner. Even though
reduction of volume is more economical and efficient. There are several different methods that are used to
reduce the volume so as to prevent loss of progenitor cells during cryopreservation (Saunders Comprehensive
Veterinary Dictionary, 2016). Such as, density gradient separation that is used for separating particles such as
DNA where the sample will be placed on a preformed gradient like sucrose or cesium chloride [7],
sedimentation of red cells by gelatin, rouleaux formation induced by hydroxyethyl starch (HES) where the
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volume of cord blood can be reduced by enhancing sedimentation of red cells by rouleaux formation that
is induced by a strong sedimenting agent like 6% hydroxyethyl starch (Hespan), centrifugation, and
differential centrifugation with expression of RCB and plasma.
Midwives who are trained in collecting cord blood, covering a period of 24 hours. At the time of delivery; sex,
weight and condition of the infant is normally learned. The cord is then being doubly clamped and transacted
within 10– 30 seconds after the delivery of newborns. To ensure sterility the free end of the cord is wiped in
betadine(Cord Blood Banking – How Baby’s Cells are Extracted & Stored? | LifeCell, 2017).
When the placenta is still inside the uterus, the umbilical vein is punctured, and cord blood is collected using
the gravity in the collection bag. If the birth of the baby is caesarean section or it is a multiple birth, the UCB
is collected from the placenta that is removed. No matter the birth type, always a maximum effort is taken to
obtain the highest possible amount of UCB. From the collected UCB, 20ml of the venous blood that was
obtained is frozen at−80°C. The other amount of UCB units that was stored in 4°C be transported, with all the
related paperwork and is processed within 24 hrs. The collected amounts of UCB units are then calculated
excluding the weight of the bags(Dı´az, 2000).
Several different tests should be run of the collected UCB; therefore, 3ml samples are taken for HLA typing,
Nucleated cell (NC) count, CD34+ cell count, Progenitor cell assays, Tests for fungal, aerobic and anaerobic
bacteriology cultures. The UCB unit will be transferred to a 150-ml bag and the HES solution will be directly
added to the collection bag under sterile conditions which has a proportion of 2:1 of washing the bag before
mixing it with the cord blood. The UCB units will be first centrifuged at 40 g for 5 minutes; because of this
the WBC (White Blood Cells)-rich supernatant is expressed by the NPBI (Non-Protein Bound Iron)Compomat
G4 system into the original collection bag. A second centrifugation will be done at 400 g for 10 minutes where
the plasma is discarded into a satellite bag. After these centrifugations, the remaining volume of UCB units
has a mean volume of 27 ml of WBC and plasma. The process is performed in a closed system with the use of
sterile connecting devices to reduce bacterial infections due to contamination. The NK (Natural Killer) and
CD34+ cell estimations are repeated using the pelleted fraction so that there is a monitoring of the quality of
the process. WBC amounts present in the collecting bag is transferred to a freezing bag(Dı´az, 2000).
Cryopreservation of the cord blood is done by cryopreservation of processed UCB units in an automated
microprocessor-controlled rate freezer. After the WBC is chilled the cold freezing cryopreservative solution
containing 60% DMSO is added drop wise for 15 minutes. Samples for quality control of cryopreservation
procedure is then extracted before freezing and cryopreserved into cryotubes with the bag. The cells will then
be immediately placed inside aluminium cassettes in the chamber of the cell freezer, that uses two
thermocouple probes placed in a sample containing the freezing solution (M-Reboredo N, 2000).
The cryopreservation methodology is by 1°C/min cooling down to −60°C, followed by a drop to −120°C,
5°C/min.
At the end of the freezing procedure the cells are stored in liquid nitrogen freezer. Immunophenotyping of the
CD34+ cell estimation iscarried out on the whole blood before starting the processing and the volume
reduction. The CD34+ cell number is calculated on the amount of the WBC units present after the
processing(Dı´az, 2000).
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3. Advantages of Cord Blood Treatment
The collecting procedure of cord blood is simple and has no medical risk to the mother or newborn baby. The
midwives and the responsible staff collects the cord blood from the placenta and then sends it for processing.
The cord blood is collected in advance therefore there is plenty of time to test it and for storage and make sure
that it is ready use when needed( Advantages and Disadvantages of Cord Blood Treatment, 2017)].
Perhaps one of the most important advantage is that cord blood transplantations do not require a perfect match
(HLA typing). Research studies have shown that cord blood transplants can be performed in cases that the
donor and the recipient are partially matched. This is because even the partially matched cord blood
transplants can be performed where cord blood increases the patient’s chance to find a more suitable donor.
An estimated number of example, that a national inventory of 150,000 cord blood units that would provide
acceptable matches for at least 80-90% of United States patients (Are there and unfavorable aspects of cord
blood? - National Cord Blood Program, 2017).
Umbilical stem cell cord blood promises to provide the solution to many critical medical conditions. Most
cord blood transplantation has been associated with lower rates of GvHD (Graft vs Host Diseases)(Are there
and unfavorable aspects of cord blood? - National Cord Blood Program, 2017).
4. Disadvantages of Cord Blood Treatment
The volume of cord blood collected is relatively small therefore the quality stem cells that are used for
transplantation much less than that in peripheral blood or bone marrow. If the average total nucleated cell dose
(number of nucleated cells per kilogram of the patient's weight) in a cord blood is less than about 1/10th of the
average bone marrow then as a consequence, the engraftment (the return of nucleated blood cells, red blood
cells and platelets) to the patient’s blood is slower with cord blood than with bone marrow transplants. This is
a major problem for adults and adolescents because they need more quantity of stem cells for transplantation
(Maslova, 2015)].
Cord blood transplantation can exposethe patient to one of the rarest genetic disorders of the immune system
or blood. This disorder is not detectable while testing the cord blood sample, but it remains (What are the
advantages of cord blood? - National Cord Blood Program, 2017).
The donor cord blood stem cells that are donated by a newborn baby are unavailable for an extra donation of
cord blood. Therefore, if by any chance the first cord blood unit fails, then a second unit should be obtained
from a different donor.
The American Academy of Pediatrics’ says that the chance of a baby needing its own stored cells is
approximately 1:1000 to 1:200,000. Therefore, private blood banking is a waste of money and resources
( Advantages and Disadvantages of Cord Blood Treatment, 2017).
5. Comparison between old and new methods of extracting and preserving umbilical cord stem cells
5.1. Early extraction methods
The collected cells were cultured in a dish and then transplanted into mice, ex vivo expansion and then in vitro
transplantation. Once the cells were grown, they were spun in a centrifuge in order to spin them down to
separate and extract them. A major limitation observed in this method was that too much plasma contents were
collected with not enough MNCs (Mono Nucleated Cells) and no reliable way to concentrate and isolate stem
cells(Hussain, 2012).
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5.2. Modern extraction methods
The modern methods derived by doing the similar steps in an automated way. The cord blood is first separated
into several layers such as a layer of RBCs (Red Blood Cells), a layer of plasma and an in between layer
which is known as the Buffy layer. Buffy layer is known to be rich with white blood cells and most essential
stem cells. Then a suitable processing method is used, to help better separation of cord blood into these
multiple layers, allowing for easier extraction of more stem cells (Hussain, 2012).
Five separation methods used are Plasma Depletion, Density Gradient, Hetastarch, PrepaCyte and Automated
centrifugal machine (Kawasaki-Oyama, 2008).
Currently the PrepaCyte is the latest and best used proprietary method. It is very similar to the closed method
but uses a machine (Basford, 2010).
6. Comparison
As it is known umbilical cord blood (UBC) is a rich source of stem cells which can be used to treat diseases.
However, earlier, this cord waste discarded as waste material as its properties were hidden. For these stem
cells to come into use the umbilical cord blood has to be collected and stored for use when needed. These cells
can easily lose their differentiable properties by external conditions if not looked into carefully (Hua, 2013).
Shortly after the delivery of the baby, the blood from the umbilical cord is collected using a syringe and
cannula and collected into a bag containing antibiotics and other necessary elements for keeping the blood safe
until it is correctly preserved. This is called the closed technique of collection since the umbilical cord is not
cut or disturbed in order to collect the blood(Hussain, 2012).
Cryopreservation techniques are then used to store the blood in blood banks public and private.
Cryopreservation is the use of extremely low temperatures maintained in order to preserve the structure of
intact living cells, along with creating a stable environment through which the cells can be preserved and
stored for future use. This method is considered most easy and reliable (Keiger, 2011).
In order to collect large number of stem cell the extraction process must be entirely or partially automated.
This closed system to its maximum extent prevents the possible happenings of bacterial contamination after
collection and before storage.
It is also needed that maximum number of nucleated cells and progenitors to enable engraftment. This was the
disadvantage seen in early separation techniques by density gradient techniques which resulted in the loss of
mononuclear cells; hence the modern methods derived which suggested that storing the cord blood before
separation. But this too is not the most ideal method as it is essential for minimum volume to be stored in cord
blood banks as only then will it be economical and efficient (Beeravolu, 2017).Yet, the modern methods are
being updated in order to reduce the volume to be stored along with reducing the loss of progenitor cells.
Some modern methods are density gradient separation; sedimentation of red cell by gelatin, rouleaux
formation induced by hydroxyethyl starch (HES) and centrifugation, and differential centrifugation with
expression of RCB and plasma (M-Reboredo N, 2000).
Finally having an optimal yield of mononuclear cells is the most important aspect of UCB collection. Along
with considering the factors which influence the amount of fetal blood which is actually remaining in the
placenta and umbilical cord after clamping and dissection (M-Reboredo N, 2000).
From this understanding it is easy to understand the advantages of modern extraction and preservation
methods. Firstly, currently the entire segment of the umbilical cord is stored, and this helps to preserve many
varied types of cells which can be used in future therapies including cell types such as mesenchymal stem
cells, endothelial cells, perivascular cell, growth factors and proteins and epithelial cells which all can be
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found in parts of the stored segment. It also processes the cord blood after preservation and this helps is
treating a wider range of treatments and able to be used for more clinical applications as the stem cells will be
more accepted in such cases. Also, cord blood is stored in over 6 vials which helps in samples being thawed
independently at different times and also allows the potential for multiple therapeutic uses (Taghizadeh, 2017).
Since UCB is known to have higher frequency of progenitor cells, and higher number of early and committed
progenitor cells, their ability to form colony forming units’ granulocyte macrophage CFU-GMO is highest.
Along with this property they also have non-hematopoietic stem cell and other cell type precursors (Basford,
2010).
Although UCB stem cells are known to have many benefits in transplantations one major disadvantage is the
low amount of total nucleated cell (TNC) number able to be collected from a single unit. Factors such as unit
size, number of previous pregnancies, age of mother, limited volumes available for collection from each
sample, and processing methods used, affect for the total collectable cell count. Hence, in order to actually
make cord blood banking a more practical option, more efficient processing method should be
considered(Basford, 2010).
7. Therapeutic uses of umbilical cord blood
Umbilical cord blood of human is a rich source of hematopoietic stem cells, totipotent cells and pluripotent
cells. These cells provide outstanding health treatments in medical industries. The stem cells in the cord blood
has the capability to develop into different type of cells so they can produce organ specific tissue in special
conditions, so cord blood is termed as regenerative medicine[6]. UCB is a good hematopoietic source and
perhaps one of the most important cells that could be derived from UCB is Natural Killer cells. These cells can
kill different targets such as cancer or virally-infected cells without any prior activation(Lubin BH, 2007)
(Roura, 2015).
Umbilical cord blood is cryopreserved for future use related to most medical conditions and disorders due to
its lifesaving properties. It is also used for developing the therapies for incurable diseases. Some of the
diseases cured are cancers and blood disorders which are considered to be fatal diseases (Moghul, 2013).
Umbilical cord blood solves many problems in the medical field as there are no counter effects after treatment.
It also helps to recover from the harmful diseases which are not cured over generations as when these genetic
disorders are cured then they do not pass on to next generation. Diseases such as Alzheimer’s, Arthritis,
Asthma, Cancer, Diabetes, Heart diseases and Strokes can be completely eradicated from a family history if
cured using umbilical cord blood. It is a prevention method from the genetic disorder being passed down to
the following generations(Moghul, 2013).
Like cord blood, the cord tissue which is termed as Wharton’s jelly is found in the umbilical cord. This
Wharton’s jelly with Poly-Vinyl Alcohol is useful for mainly treating the skin wounds for humans
(Mothersofchange.com, 2012). There is so much difficulty and failure when therapies are used for healing the
chronic skin wounds. Examples for some conditions that treated with stem cells are mentioned in the Table 1
(Waller-Wise, 2011).
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Table 1: Examples of Conditions Treated with Stem cell Transplants. The table provides a brief understanding
about different types of cancers, blood disorder, congenital metabolic disorders and immunodeficiencies
which can be treated using stem cell transplants.
Cancers Blood disorders Congenital metabolic
disorders
Immunodeficiencies
Acute lymphocytic
leukemia
Sickle-cell anemia Adrenoleukodystrophy Adenosine deaminase
deficiency
Acute myelogenous
leukemia
Fanconi’s anemia Gunther’s disease Wiskott-Aldrich’s
syndrome
Chronic myelogenous
leukemia
Thalassemia Gaucher’s disease Duncan’s disease
Myelodysplastic
syndrome
Evan’s syndrome Hurler’s syndrome Ataxia-telangiectasia
Neuroblastoma Congenital cytopeni
1
a Hunter’s syndrome DiGeorge’s syndrome
Hodgkin’s disease Aplastic anemia Krabbe’s disease Myelokathexis
Non–Hodgkin’s
lymphoma
Diamond–
Blackfananemia
Sanfilippo’s syndrome Hypogammaglobuline
mia
Burkitt’s lymphoma Amegakaryocytic
thrombocytopenia
Tay-Sachs’ disease SevereCombined
immunodeficiency
Wharton’s jelly serves as a good source to heal the wounds as they have the properties like high plasticity,
proliferative, differentiation capability, and also low immunogenicity.
This umbilical cord blood has the capacity to differentiate into blood due to the composition of increased
count of hemoglobin, hematocrit, leukocytes, reticulocytes, and nucleated (immature) red blood cells with
presence of immature white blood cells (G.H. Mamoury, 2003). Therefore, that the cord blood collected from
the newborn infants is used as an alternative to bone marrow in transplantations and to restore immunological
dysfunctions (Waller-Wise, 2011).
The hematopoietic stem cell is the main composition of the cord blood which has the capability to differentiate
into blood and also, they are known to have the antigens CD133, CD34 and CD45 they can be induced during
in vitro differentiate in too many linages such as erythroid, megakaryocytic and monocyticetc (Roura, 2015).
Mesenchymal stem cells derived from cord blood has high morphological and also molecular similarities
when compared to mesenchymal stem cells derived from the bone marrow. They are being used in medical
industry due to their potential of rapid multiplication (Roura, 2015). The mesenchymal cell is used for
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growing in popularity due to its properties like immunomodulatory, anti-inflammatory and tissue regenerative
properties (Lubin BH, 2007).
Neurons cannot be regenerated but mesenchymal stem cells make it possible where it can regenerate or
replace the damaged neurons and also increases the myelination of axons. It mainly helps to reduce apoptotic
cell death by maintaining homeostasis. During transplantation people suffer due to rejection of the
transplanted organ by the body so this MSCs controls the immune cells and prevents the inflammation and
rejection caused due to transplantation. MSCs can also replace and repair the blood vessels so they are termed
as potential therapeutic remedy for the patients suffering from stroke (Hamad Ali, 2012).
Multipotent non-hematopoietic stem cellsare the final type of stem cells found in the cord blood. These cells
can differentiate in cell surface markers and various other cells which represent the three germ layer cells
(Roura, 2015).
The umbilical cord blood banking is used mainly in order to treat the genetic diseases caused which cannot be
cured by the normal treatments. Many diseases are treated using the cord blood stem cells because these cells
have the potential to transform into the differentiated cells needed by treating these cells in specialized cell
culture medium so that, it can even develop into a specific organ tissue needed. This is the main reason for the
banking of the umbilical cord blood (Hua, 2013).
When there is a disease in the first-born baby then the cord blood obtained from the birth of next baby can
help to overcome the disease likewise that cord blood obtained can be also used for the same child if there is
any inherited genetic disorder, so the banking of the umbilical cord blood plays an important role. As this cell
has the potential to differentiate into non-hematopoietic tissue such as cardiac, neurologic, pancreatic and skin
tissues due to the presence of pluripotent stem cells so it can be used to treat the diseases like bone marrow
failure, hemoglobinopathies, immunodeficiency, and also the inborn errors of metabolism. This is the main
therapeutic use of UCB banking in the clinical or medical field (B. Anthony Armson, 2015).
There is both public and private banking to preserve or to store the umbilical cord blood. Mostly public bank
is recommended by the clinicians as there is no fee required for the public cord blood banking and it does not
work for any profit. Also, unrelated transplantations are possible to help the people who are actually in need of
it. Public cord blood bank is considered to be easily accessed by the public and also involves direct donations
of cord blood (CryoCell International, 2017) (Virginia P. Studdert, 2011).
Even though private bank is not mostly recommended it also plays an important role in some aspects. It can be
mainly accessed only for family use also it is suggested as a good choice to get rid of hereditary diseases. This
is not easily assessable for public so it can be preserved for related transplantations because only the donor has
all the rights on their cord blood unit (Wharton’s Jelly: Miracle Tissue, 2017).
8. Discussion
The umbilical cord blood was discarded as a waste material but today it is considered to be a regenerative
medicine in order to produce the organ tissues (Hua, 2013) This cord blood collected from the umbilical cord
is composed of mesenchymal stem cells, hematopoietic stem cells and also multipotent non-hematopoietic
stem cells which has therapeutic uses as these stem cells are used to treat cancers, blood disorders, congenital
metabolic disorders and immunodeficiencies (Roura, 2015). Cord blood also contains some non-hematopoietic
stem cells like EPCs (endothelial progenitor cells), MSC (Mesenchymal Stromal Cells), USSC (Unrestricted
Somatic Stem Cells),VSEL (Very Small Embryonic-Like stem cells), MLPC (Multi Lineage Progenitor
Cells), and neuronal progenitor cells. Mostly the transplantations of bone marrow require surgery to obtain
from the donor and also causes rejection but when the transplantation is performed with umbilical cord blood
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then it prevents the rejection of transplanted organ and also no need of surgery to obtain the cord blood
(Moghul, 2013).
Like umbilical cord blood Wharton’s jelly is also derived from the umbilical cord blood which is a rich source
of mesenchymal stem cells. Wharton’s jelly is a gelatinous tissue within the umbilical cord which forms the
umbilical cord matrix in order to cover the veins and arteries of umbilical cord. This Wharton’s jelly is used in
healing of chronic injuries (Zoe, 2016) Cryopreservation is followed to preserve cord blood and Wharton’s
jelly as it is an easy and reliable method (Basford, 2010).
The collection of the cord blood is the important process in the banking of umbilical cord blood which takes
only some minutes. The collection of the cord blood is done during the delivery by clamping the umbilical
cord. Once when the umbilical cord is clamped it is wiped with an antiseptic and then a needle is inserted in
order to obtain 60 ml of the cord blood early clamping should not be followed as it leads to anemia (Pegg,
2007).
There are two different types of umbilical cord blood bank banks which is very familiar they are public cord
blood bank and private cord blood bank. There is also another one as direct donation cord blood bank
(Wharton’s Jelly: Miracle Tissue, 2017).
The cord blood has a small chance to be used by the donor so it is better to store it in a public bank as it can
save a life. In addition, many legal and ethical aspects must be considered in private cord blood banking. It is
said to be a biological insurance stored for the future family use as it is regarded as the once in a life time
opportunity many of them chose this option(Buatovich, 2016).
USA pioneered in the umbilical cord blood banking. USA has twenty-eight Public cord blood banks and
twenty-nine private cord blood banks (Disadvantages of Cord Blood Banking, 2017). UK also implement both
Private and Public Cord Blood banking and there are two public Cord blood banks and six Private cord blood
banks in UK. There are no any Public cord blood banks in the Egypt all of them are private cord blood banks,
there are five private cord blood banks in Egypt which are established by other countries such as UK,
Switzerland and India. Whereas there is an Egyptian private cord blood bank named as cell safe bank
(Disadvantages of Cord Blood Banking, 2017). South Africa being prominent country in African continent,
has three private UCB Banks.Singapore a well-developed country in Asian continent has only one public cord
blood banks. India, as a developing country plays a major role in cord blood banking. India has four public
cord blood banks and fourteen private cord blood banks (Seah, 2014). Sri Lanka is about to establish a Cord
Blood Bank. This will be available at National Blood Centre under the guidance of Dr.
VijithGunasekara(Hettiarachchi, 2012).
Only 0.04% of the stem cell used by the same donor and only 0.07% is used for the familial usage such as for
siblings. Even though this reduces 50% of the risk of graft versus host diseases private bank cost additional $
1,374,246 for each year [24]. More than 600000 UCB units have been stored in world wide. But only >30000
transplants have been performed. Even though there are more than 20 million Adult volunteer donors
registered in National Marrow Donor program (Karen K. Ballen, 2013).
Acknowledgements
International College of Business and Technology (ICBT) No. 36, De Kretser Place, Bambalapitiya, Colombo
04, Sri Lanka and the Department of Biomedical Science at ICBT is gratefully acknowledged for all the
Support and Encouragement.
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References
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... 7 Research studies have shown that cord blood transplants can also be performed in cases even when the donor and the recipient are partially matched, so cord blood increases the patient's chance to find a more suitable donor. 8,9 However, with increasing differences in the donor and recipient HLA systems, the risk of delayed or absent hematopoietic reconstitution also increases in UCB transplantation. 8,10 In contrast, the greater availability of high-quality and high-cell-content UCB units has resulted in increasingly improved engraftment and survival outcomes after UCB transplantation. ...
... 8,9 However, with increasing differences in the donor and recipient HLA systems, the risk of delayed or absent hematopoietic reconstitution also increases in UCB transplantation. 8,10 In contrast, the greater availability of high-quality and high-cell-content UCB units has resulted in increasingly improved engraftment and survival outcomes after UCB transplantation. 11,12 There are various providers worldwide, which accompany or sometimes even offer the process from collection to storage. ...
... 16 Using sterile techniques and a closed system during the whole preparation process is usually used to reduce the risk of any bacterial contamination. 8,16 Freezing bags or tubes must comply with the current state of technology and exclude cross-contamination between different samples during storage. 16 The aim of this study was to investigate the bacterial contamination rate related to the delivery mode including collection experiences of the staff and collection sample volume. ...
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Cord blood (CB) collected at birth has become a valuable stem cell source for hematopoietic stem cell transplantation (HSCT). However, the collection of umbilical cord blood always bears a risk of microbiological contamination, both in vaginal birth and in cesarean section. A total of 10 054 umbilical cord stem cell samples were successfully cryopreserved between 2010 and 2020, of which 783 (8%) samples were tested positive for bacterial contamination. Umbilical CB with a volume of less than 60 mL showed a bacterial contamination rate of 12%, and above 60 mL volume a rate of 6% was found demonstrating an inverse relationship between sample volume and contamination rate (correlation coefficient r = -0.9). The contamination rate was associated with the mode of delivery and showed a significantly higher contamination rate of 9.7% when compared with cesarean deliveries (1.4%). The 10-year period consistently shows an average contamination rate between 4% and 6% per year. It is conceivable that the inverse relationship between volume and contamination rate might be related to thinner veins although no scientific evidence has been provided so far. The lower contamination rate in cesarean sections appears to be related to the sterile operating setting. Overall, the rate of bacterial contamination varies and depends on the type of birth, the way of delivery, and probably the experience of the staff.
... Using sterile Pasteur pipette, 10 μl of cell trypan blue suspension mixture was transferred to a hemocytometer chamber and covered with a coverslip firmly in place, viable cells (uncolored cells) in each of the four corner squares on either side of the center chamber were calculated, average the counts multiplied by 2×10 4 to give the number of cells/ml. 19 ...
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Aflatoxins (AFs) are secondary metabolites having a high cytotoxic potential and have an active carcinogenic compound. This study aimed to isolate hematopoietic stem cells (Precursors of mononuclear stem cells) from human umbilical cord blood. Collection of umbilical cord blood samples from cord blood was accomplished immediately after delivery, while the placenta is intrauterine by used closed system method. The mononuclear cells (MNCs) were obtained from umbilical cord blood by using the Buffy coat method. The previous cells have been Hematopoietic Stem Cells (HSCs) used in the cytotoxicity test of AF extract. The mononuclear cells were cultured in DMEM medium with 10% FCS. An experimental study was conducted in vitro on these cells to confirm the toxic effects of AF with different concentrations of AFs. The results showed that the inhibition rate increases seriously with the increase of AF concentration. Aspergillus flavus in peanut produces the highest concentration of AF, while in the soil, contamination with crude oil A. flavus produces the least concentration of AF. Additionally, the concentration of 1 μg /ml of AF extracted from the aflatoxigenic isolates which can kill 100% of hematopoietic stem cells.
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The human umbilical cord (UC) and placenta are non-invasive, primitive and abundant sources of mesenchymal stromal cells (MSCs) that have increasingly gained attention because they do not pose any ethical or moral concerns. Current methods to isolate MSCs from UC yield low amounts of cells with variable proliferation potentials. Since UC is an anatomically-complex organ, differences in MSC properties may be due to the differences in the anatomical regions of their isolation. In this study, we first dissected the cord/placenta samples into three discrete anatomical regions: UC, cord-placenta junction (CPJ), and fetal placenta (FP). Second, two distinct zones, cord lining (CL) and Wharton's jelly (WJ), were separated. The explant culture technique was then used to isolate cells from the four sources. The time required for the primary culture of cells from the explants varied depending on the source of the tissue. Outgrowth of the cells occurred within 3 - 4 days of the CPJ explants, whereas growth was observed after 7 - 10 days and 11 - 14 days from CL/WJ and FP explants, respectively. The isolated cells were adherent to plastic and displayed fibroblastoid morphology and surface markers, such as CD29, CD44, CD73, CD90, and CD105, similarly to bone marrow (BM)-derived MSCs. However, the colony-forming efficiency of the cells varied, with CPJ-MSCs and WJ-MSCs showing higher efficiency than BM-MSCs. MSCs from all four sources differentiated into adipogenic, chondrogenic, and osteogenic lineages, indicating that they were multipotent. CPJ-MSCs differentiated more efficiently in comparison to other MSC sources. These results suggest that the CPJ is the most potent anatomical region and yields a higher number of cells, with greater proliferation and self-renewal capacities in vitro. In conclusion, the comparative analysis of the MSCs from the four sources indicated that CPJ is a more promising source of MSCs for cell therapy, regenerative medicine, and tissue engineering.
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In light of pioneering findings in the 1980s and an estimation of more than 130 million global annual births, umbilical cord blood (UCB) is considered to be the most plentiful reservoir of cells and to have regenerative potential for many clinical applications. Although UCB is used mainly against blood disorders, the spectrum of diseases for which it provides effective therapy has been expanded to include non-hematopoietic conditions; UCB has also been used as source for regenerative cell therapy and immune modulation. Thus, collection and banking of UCB-derived cells have become a popular option. However, there are questions regarding the cost versus the benefits of UCB banking, and it also raises complex ethical and legal issues. This review discusses many issues surrounding the conservation of UCB-derived cells and the great potential and current clinical applications of UCB in an era of new therapies. In particular, we describe the practical issues inherent in UCB collection, processing, and long-term storage as well as the different types of 'stem' or progenitor cells circulating in UCB and their uses in multiple clinical settings. Given these considerations, the trend toward UCB will continue to provide growing assistance to health care worldwide.
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In an era increasingly focused on quality improvement and cost containment, more emphasis is being placed on wiser utilization of medical care resources. One underutilized resource in early neonatal care is umbilical cord blood. Umbilical cord blood can be utilized for admission laboratory studies in neonates thereby avoiding a significant phlebotomy event in the first minutes to hours of life. Additionally, umbilical cord blood can also be safely “transfused” into the neonate via delayed cord clamping or milking of the umbilical cord. This has been demonstrated to be particularly beneficial in premature infants by decreasing the rate of intraventricular hemorrhage. Delayed cord clamping has been formally endorsed by a number of medical societies, however it has not yet been universally adopted by obstetricians and neonatologists. Both uses of umbilical cord blood for neonatal admission laboratory testing and delayed cord clamping/milking of the umbilical cord have resulted in decreased transfusion rates as well as other outcomes reviewed herein.
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Although the characteristics of SC, including UC-derived cells, are a dramatically discussed issue, this review will focus particularly on some controversial issues regarding clinical utility of cells isolated from UC tissue. UC-derived cells have several advantages compared to other types and sources of stem cells. The impact of UC topography on cell characteristics is briefly discussed. The necessity to adapt existing methods of cell isolation and culturing to GMP conditions is mentioned, as well as possible cryopreservation of this material. Light is shed on some future perspectives for UC-derived cells.
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To date, various types of cells for seeding regenerative scaffolds have been used for bone tissue engineering. Among seed cells, the mesenchymal stem cells derived from human umbilical cord Wharton's jelly (hUCMSCs) represent a promising candidate and hold potential for bone tissue engineering due to the the lack of ethical controversies, accessibility, sourced by non-invasive procedures for donors, a reduced risk of contamination, osteogenic differentiation capacities, and higher immunomodulatory capacity. However, the current culture methods are somewhat complicated and inefficient and often fail to make the best use of the umbilical cord (UC) tissues. Moreover, these culture processes cannot be performed on a large scale and under strict quality control. As a result, only a small quantity of cells can be harvested using the current culture methods. To solve these problems, we designed and evaluated an UC Wharton's jelly repeated culture device. Using this device, hUCMSCs were obtained from the repeated cultures and their quantities and biological characteristics were compared. We found that using our culture device, which retained all tissue blocks on the bottom of the dish, the total number of obtained cells increased 15-20 times, and the time required for the primary passage was reduced. Moreover, cells harvested from the repeated cultures exhibited no significant difference in their immunophenotype, potential for multilineage differentiation, or proliferative, osteoinductive capacities, and final osteogenesis. The application of the repeated culture frame (RCF) not only made full use of the Wharton's jelly but also simplified and specified the culture process, and thus, the culture efficiency was significantly improved. In summary, abundant hUCMSCs of dependable quality can be acquired using the RCF.
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This policy statement is intended to provide information to guide pediatricians, obstetricians, and other medical specialists and health care providers in responding to parents' questions about cord blood donation and banking as well as the types (public versus private) and quality of cord blood banks. Cord blood is an excellent source of stem cells for hematopoietic stem cell transplantation in children with some fatal diseases. Cord blood transplantation offers another method of definitive therapy for infants, children, and adults with certain hematologic malignancies, hemoglobinopathies, severe forms of T-lymphocyte and other immunodeficiencies, and metabolic diseases. The development of universal screening for severe immunodeficiency assay in a growing number of states is likely to increase the number of cord blood transplants. Both public and private cord blood banks worldwide hold hundreds of thousands of cord blood units designated for the treatment of fatal or debilitating illnesses. The procurement, characterization, and cryopreservation of cord blood is free for families who choose public banking. However, the family cost for private banking is significant and not covered by insurance, and the unit may never be used. Quality-assessment reviews by several national and international accrediting bodies show private cord blood banks to be underused for treatment, less regulated for quality control, and more expensive for the family than public cord blood banks. There is an unquestionable need to study the use of cord blood banking to make new and important alternative means of reconstituting the hematopoietic blood system in patients with malignancies and blood disorders and possibly regenerating tissue systems in the future. Recommendations regarding appropriate ethical and operational standards (including informed consent policies, financial disclosures, and conflict-of-interest policies) are provided for physicians, institutions, and organizations that operate or have a relationship with cord blood banking programs. The information on all aspects of cord blood banking gathered in this policy statement will facilitate parental choice for public or private cord blood banking.
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Cryopreservation is the use of very low temperatures to preserve structurally intact living cells and tissues. Unprotected freezing is normally lethal and this chapter seeks to analyze some of the mechanisms involved and to show how cooling can be used to produce stable conditions that preserve life. The biological effects of cooling are dominated by the freezing of water, which results in the concentration of the solutes that are dissolved in the remaining liquid phase. Rival theories of freezing injury have envisaged either that ice crystals pierce or tease apart the cells, destroying them by direct mechanical action, or that damage is from secondary effects via changes in the composition of the liquid phase. Cryoprotectants, simply by increasing the total concentration of all solutes in the system, reduce the amount of ice formed at any given temperature; but to be biologically acceptable they must be able to penetrate into the cells and have low toxicity. Many compounds have such properties, including glycerol, dimethyl sulfoxide, ethanediol, and propanediol.
Article
DOI: http://dx.doi.org/10.5915/42-1-5197 Umbilical cord blood (UCB) is a very valuable source of both hematopoietic and pluripotent stem cells. It is a readily available, easily collected source that does not cause inconvenience or harm to the donor. It has been shown to be useful in the treatment of several neoplastic and non-neoplastic serious diseases. UCB transplants have been proven to be life saving in many of these conditions. It results in better survival rates and less GVHD than the traditional treatment with bone marrow transplants. It probably can fill the gap that is caused by the lack of a suitable bone marrow match. There is no ethical or moral objection to its use. UCB donation should be encouraged. UCB can be collected and cryopreserved for at least 15 years in blood banks, which can be public and private. The pros and cons of each type are described. The family, when donating cord blood, has to choose which type it prefers. Detailed and balanced information should be given to the parents by their healthcare providers before they make that decision. Health authorities should support the development of the public banks because by their nature they do not make money and they provide the much needed help for those who cannot afford private banking. Ethical concerns including proper informed consent, linkage of the donor to the donated units, truth in advertising by the private banks and distributive justice are discussed. Also discussed is the question of the appropriateness of selective conception of a baby to be a potential donor.to come