ArticlePDF Available

Fetal bovine serum (FBS): Past – present – future

Authors:
J.B.F. Van der Valk at Utrecht University
J.B.F. Van der Valk
Karen Bieback at Universität Heidelberg
Karen Bieback
Christiane Buta
Christiane Buta
Christiane Buta
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Brett Cochrane
Brett Cochrane
Brett Cochrane
  • This person is not on ResearchGate, or hasn't claimed this research yet.
Show all 17 authorsHide
Download full-text PDFDownload full-text PDFDownload full-text PDF
Read full-text
Download citation

Abstract and Figures

The supplementation of culture medium with fetal bovine serum (FBS, also referred to as 'fetal calf serum') is still common practice in cell culture applications. Due to a number of disadvantages in terms of quality and reproducibility of in vitro data, animal welfare concerns, and in light of recent cases of fraudulent marketing, the search for alternatives and the development of serum-free medium formulations gained global attention. Here, we report on the 3rd Workshop on FBS, Serum Alternatives and Serum-free Media, where (a) regulatory aspects, (b) the serum dilemma, (c) alternatives to FBS, (d) case-studies of serum-free in vitro applications, and (e) the establishment of serum-free databases, were discussed. The whole process of obtaining blood from a living calf fetus to using the FBS produced from it for scientific purposes is de facto not yet legally regulated, despite the existing EU-Directive 2010/63/EU on the use of animals for scientific purposes. Together with above mentioned challenges, several strategies have been developed to reduce or replace FBS in cell culture media in terms of the 3Rs (Refinement, Reduction, Replacement). Most recently, releasates of activated human donor thrombocytes (human platelet lysates) have been shown to be one of the most promising serum alternatives when chemically defined media are not yet an option. Additionally, new developments in cell-based assay techniques, advanced organ-on-chip and microphysiological systems are covered in this report. Chemically-defined serum-free media are shown to be the ultimate goal for the majority of culture systems, and examples are discussed.
Scanning electron micrograph of resting (left) and activated (right) human thrombocytes, immobilized on sterile filters When platelets are activated, they undergo characteristic morphological changes and form long pseudopodia to adhere to the sites of vessel wall injury. In addition, thrombocytic granule factors are released by exocytosis: vasoactive peptides, factors for initiating and maintaining the clotting cascade, and growth factors for subsequent wound healing. Filters from Millipore Inc., pore size 0.22 µm. Micrograph taken by G. Gstraunthaler.
Scanning electron micrograph of resting (left) and activated (right) human thrombocytes, immobilized on sterile filters When platelets are activated, they undergo characteristic morphological changes and form long pseudopodia to adhere to the sites of vessel wall injury. In addition, thrombocytic granule factors are released by exocytosis: vasoactive peptides, factors for initiating and maintaining the clotting cascade, and growth factors for subsequent wound healing. Filters from Millipore Inc., pore size 0.22 µm. Micrograph taken by G. Gstraunthaler.
… 
Test for the eye irritation potential of seven dilutions of sodium dodecyl sulphate toward L929 fibroblasts A: extracellular acidification rate (EAR), B: modulus of impedance (ІZІ)). The first five recordings of the basal rates are shown, followed by seven dilutions of sodium dodecyl sulphate.
Test for the eye irritation potential of seven dilutions of sodium dodecyl sulphate toward L929 fibroblasts A: extracellular acidification rate (EAR), B: modulus of impedance (ІZІ)). The first five recordings of the basal rates are shown, followed by seven dilutions of sodium dodecyl sulphate.
… 
Test for the eye irritation potential of seven dilutions of sodium dodecyl sulfate toward L929 fibroblasts Left: extracellular acidification rate (EAR), right: modulus of impedance (IZI). The first five recordings of the basal rates are shown, followed by seven dilutions of sodium dodecyl sulfate.
Test for the eye irritation potential of seven dilutions of sodium dodecyl sulfate toward L929 fibroblasts Left: extracellular acidification rate (EAR), right: modulus of impedance (IZI). The first five recordings of the basal rates are shown, followed by seven dilutions of sodium dodecyl sulfate.
… 
Schematic view of a 4-organ microfluidic platform (a) Schematic view of the microfluidic platform showing the different cell compartments of a 4-organ system. The system contained two holders for the separate culture devices. Total fluid volume was approximately 4 mL between the chambers and reservoirs. The sizes of the culture compartments were 35.8 x 18.4 x 0.3 mm for Chambers 1, 2, 3 and 29.8 x 15.4 x 0.7 mm for Chambers 4 and 5. The connecting channel dimensions were 5.7 x 1 x 0.3 mm. (b) Shear stress distribution in each compartment of the system. Reproduced with permission from (Oleaga et al., 2016).
Schematic view of a 4-organ microfluidic platform (a) Schematic view of the microfluidic platform showing the different cell compartments of a 4-organ system. The system contained two holders for the separate culture devices. Total fluid volume was approximately 4 mL between the chambers and reservoirs. The sizes of the culture compartments were 35.8 x 18.4 x 0.3 mm for Chambers 1, 2, 3 and 29.8 x 15.4 x 0.7 mm for Chambers 4 and 5. The connecting channel dimensions were 5.7 x 1 x 0.3 mm. (b) Shear stress distribution in each compartment of the system. Reproduced with permission from (Oleaga et al., 2016).
… 
Figures - uploaded by the authors
Content may be subject to copyright.
ALTEX 35_99_20
18.pdf
Content uploaded by Gerhard Gstraunthaler
Author content
All content in this area was uploaded by Gerhard Gstraunthaler on Jun 22, 2018
Content may be subject to copyright.
ALTEX 35_99_20
18.pdf
Content uploaded by Gerhard Gstraunthaler
Author content
All content in this area was uploaded by Gerhard Gstraunthaler on May 24, 2018
Content may be subject to copyright.
vanderValk_of_171013_
v3.pdf
Content uploaded by J.B.F. Van der Valk
Author content
All content in this area was uploaded by J.B.F. Van der Valk on Oct 23, 2017
Content may be subject to copyright.
A preview of the PDF is not available

Supplementary resources (2)

ALTEX 35 99 2018
Data
January 2018
J.B.F. Van der Valk · Karen Bieback · Christiane Buta · Brett Cochrane · Gerhard Gstraunthaler
Fetal Bovine Serum (FBS): Past - Present - Future; ALTEX 35(1): 99-118 (2018)
Data
January 2018
J.B.F. Van der Valk · Karen Bieback · Christiane Buta · Brett Cochrane · Gerhard Gstraunthaler
... Serum contains numerous growth factors (e.g., fibroblast growth factor and platelet-derived growth factor) that promote fibroblast growth and inhibit epithelial cell growth [35,36]. Moreover, serum introduces variability, cost, and ethical concerns [37,38]. Contaminated serum may compromise cell culture experiments. ...
Establishment and characterization of the first immortalized vocal cord leukoplakia epithelial cell line
Article
Full-text available
  • Nov 2024
The importance of vocal cord leukoplakia (VCL) in the etiology and progression of laryngeal carcinoma has gained increasing recognition. However, research into the mechanisms of laryngeal precancerous lesions such as VCL, has been hampered by the small size of VCL epithelial cells and their limited culture lifespan. In this study, we enhanced the primary culture protocol for VCL epithelial cells and introduced simian virus 40 Large T to establish an immortalized cell line, designated hVCL-MSDEP01. We confirmed that hVCL-MSDEP01 expresses epithelial-specific genes and proteins; it also demonstrates distinct cell cycle dynamics and apoptosis rates compared with primary cells. In conclusion, hVCL-MSDEP01 serves as an ideal in vitro model for studying VCL. This cell line will substantially advance research into the etiology and progression of laryngeal carcinoma.
View
... Additionally, extensive discussions have underscored the presence of contaminants in FBS, such as non-human N-glycolylneuraminic acid, possibly impeding the translation of pre-clinical findings into clinical applications. [16][17][18] Furthermore, studies have reported that bovine collagen has the potential to elicit an immune response in humans. 19,20 Despite less scientific scrutiny in comparison to FBS concerning the use of bovine collagen Figure 11. ...
Advancing skin model development: A focus on a self-assembled, induced pluripotent stem cell-derived, xeno-free approach
Article
Full-text available
  • Nov 2024
The demand for skin models as alternatives to animal testing has grown due to ethical concerns and the need for accurate substance evaluation. These alternatives, known as New Approach Methodologies (NAMs), are increasingly used for regulatory decisions. Current skin models from primary human cells often rely on bovine collagen, raising ethical issues. This study explores self-assembled skin models (SASM) as a new method, utilizing hair follicle-derived keratinocytes reprogrammed into induced pluripotent stem cells (iPSC) and differentiated into fibroblasts and keratinocytes. The model relies on the ability of fibroblasts to secrete collagen to produce a xeno-free dermal layer and on the differentiation of keratinocytes to create a functional epidermal layer. These layers exhibited confirmed metabolic activity and the capability to withstand test substances. The successful development of SASM underscores the significance of accurate alternatives in dermatological research, providing an ethical and reliable option for substance evaluation and regulatory testing.
View
... TEER assay measurement of a biological test system involves several steps, including transporting the test system from the incubator to a biological safety cabinet or bench top, rinsing the test system apically and basolaterally with appropriate solution (see Step 2), adding the electrolyte solution(s) to the apical and basolateral sides, performing the measurement, replacing the solution with fresh culture medium (if applicable), and putting the test system back in the incubator. The measured TEER assay values can differ between biological test systems based on the cell types, properties of cell culture inserts (material type, coating, dimensions, pore size, and pore density) on which they are cultured, and reagents (e.g., medium and supplements) used (Barosova et al. 2021;van der Valk et al. 2018;Leung et al. 2020;Vigh et al. 2021;Karakocak et al. 2023;Srinivasan et al. 2015;Meindl et al. 2023). Therefore, the type of biological system, the cell types, and the membrane properties must be reported (Step 3b). ...
Minimum information for reporting on the TEER (trans-epithelial/endothelial electrical resistance) assay (MIRTA)
Article
Full-text available
  • Oct 2024
  • ARCH TOXICOL
Standard information reporting helps to ensure that assay conditions and data are consistently reported and to facilitate inter-laboratory comparisons. Here, we present recommendations on minimum information for reporting on the TEER (trans-epithelial/endothelial electrical resistance) assay (MIRTA). The TEER assay is extensively used to evaluate the health of an epithelial/endothelial cell culture model and as an indicator of the potential toxicity of a test substance. This publication is the result of an international collaboration─called the RespTox (Respiratory Toxicity) Collaborative─through which twelve laboratories shared their protocols for assessing the barrier function of respiratory epithelial cells using the TEER assay following exposure to substances. The protocols from each laboratory were reviewed to identify general steps for performing the TEER assay, interlaboratory differences between steps, the rationale for differences, whether these differences impact results or cross-laboratory comparisons between TEER measurements. While the MIRTA recommendations are focused on respiratory epithelial cell systems, these recommendations can be adapted for other cell systems that form barriers. The use of these recommendations will support data transparency and reproducibility, reduce challenges in data interpretation, enable cross-laboratory comparisons, help assess study quality, and facilitate the incorporation of the TEER assay into national and international testing guidance.
View
... The quality of FBS must be carefully controlled to prevent the spread of zoonoses like Mycoplasma infections, brucellosis, and prion diseases, ensuring a pathogen-free final product [24,35,44]. Despite its widespread use, FBS has several disadvantages, including high cost, batch-to-batch quality variation, composition variability affecting growth and productivity, contamination risks, potential toxicity at high concentrations, and ethical concerns [42,45]. Additionally, FBS is not produced in many countries, particularly in the developing regions where leishmaniasis is most prevalent and much of the research occurs [42]. ...
An Overview of Leishmania In Vitro Cultivation and Implications for Antileishmanial Screenings against Promastigotes
Article
Full-text available
  • Oct 2024
The quest for new drug candidates targeting neglected parasitic diseases has become increasingly urgent over the past decades. Advancements in formulating and optimizing drug delivery systems begin with basic research, including direct assays to evaluate the activity of molecules against parasitic stages maintained in laboratories; i.e., promastigotes. In the context of leishmaniasis, an endemic disease worldwide, the cultivation of Leishmania parasites can vary significantly across different laboratories. Factors such as culture media composition, pH, supplementation, and temperature can lead to varied drug responses in in vitro activity assays. This study aims to compile the parameters used in Leishmania spp. promastigotes cultivation protocols described in scientific articles published in indexed journals over the past ten years. The data reveal a lack of uniformity among Leishmania culture protocols, suggesting a potential bottleneck in comparing the leishmanicidal potential of in vitro drug candidates reported by different research groups. This condition is crucial to consider, because viability/inhibition assays should begin with fully-grown, healthy promastigote cultures capable of homogeneous division, thereby producing more reproducible results.
View
... This is crucial for cells to survive the stress of freezing and thawing. Previous findings by Fugisawa et al. (2019) also reported that FBS helps balance osmotic pressure during freezing and reduces ice crystal damage [38]. of the earliest and most commonly used agents in cryopreservation, especially in fields involving mesenchymal cells [39]. According to Duarte Rojas JM et al. (2024), cryopreservation and thawing of cells is more effective in either platelet lysate serum or FBS [37]. ...
Optimisation of cryopreservation conditions, including storage duration and revival methods, for the viability of human primary cells
Article
Full-text available
  • Sep 2024
Background Cryopreservation is a crucial procedure for safeguarding cells or other biological constructs, showcasing considerable potential for applications in tissue engineering and regenerative medicine. Aims This study aimed to evaluate the effectiveness of different cryopreservation conditions on human cells viability. Methods A set of cryopreserved data from Department of Tissue Engineering and Regenerative Medicine (DTERM) cell bank were analyse for cells attachment after 24 h being revived. The revived cells were analysed based on different cryopreservation conditions which includes cell types (skin keratinocytes and fibroblasts, respiratory epithelial, bone marrow mesenchymal stem cell (MSC); cryo mediums (FBS + 10% DMSO; commercial medium); storage durations (0 to > 24 months) and locations (tank 1–2; box 1–5), and revival methods (direct; indirect methods). Human dermal fibroblasts (HDF) were then cultured, cryopreserved in different cryo mediums (HPL + 10% DMSO; FBS + 10% DMSO; Cryostor) and stored for 1 and 3 months. The HDFs were revived using either direct or indirect method and cell number, viability and protein expression analysis were compared. Results In the analysis cell cryopreserved data; fibroblast cells; FBS + 10% DMSO cryo medium; storage duration of 0–6 months; direct cell revival; storage in vapor phase of cryo tank; had the highest number of vials with optimal cell attachment after 24 h revived. HDFs cryopreserved in FBS + 10% DMSO for 1 and 3 months with both revival methods, showed optimal live cell numbers and viability above 80%, higher than other cryo medium groups. Morphologically, the fibroblasts were able to retain their phenotype with positive expression of Ki67 and Col-1. HDFs cryopreserved in FBS + 10% DMSO at 3 months showed significantly higher expression of Ki67 (97.3% ± 4.62) with the indirect revival method, while Col-1 expression (100%) was significantly higher at both 1 and 3 months compared to other groups. Conclusion In conclusion, fibroblasts were able to retain their characteristics after various cryopreservation conditions with a slight decrease in viability that may be due to the thermal-cycling effect. However, further investigation on the longer cryopreservation periods should be conducted for other types of cells and cryo mediums to achieve optimal cryopreservation outcomes.
View
Use of serum-free media for peripheral blood mononuclear cell culture and the impact on T and B cell readouts
Article
  • Nov 2024
Introduction As part of a wider programme of work developing next-generation risk assessment approaches (NGRA) using non-animal methods (NAMs) for safety assessment of materials, Unilever SEAC is exploring the use of a peripheral blood mononuclear cell (PBMC) system to investigate how cells from different arms of the human immune system are impacted by different treatments. To maximise human relevance, the cell cultures are supported by human serum, but this came with some challenges, including an inability to measure induced levels of immunoglobulins due to high background levels. Therefore, a study comparing use of human sera containing media with three different chemically defined serum-free media was undertaken. Materials and Methods PBMC were isolated from healthy donors and cultured in the absence (media alone) or presence of stimulation reagents (CpG-ODN plus IL-15, Pokeweed Mitogen (PWM) or Cytostim (CS)), in RPMI plus human serum, AIM-V, CTS OpTmizer T cell expansion SFM or X-VIVO 15 media. T cell (CD4⁺ and CD8⁺) and B cell proliferation and viability were measured after 6 days, along with levels of total IgG in the cell culture supernatants. Results Each of the serum-free media tested supported good levels of viable and proliferating T cells and B cells over the 6 days of culture, with only a few, small differences across the media, when there was no stimulation. They also enabled detection of a stimulation-evoked increase in IgG levels. There were however some differences in the viability and proliferation responses of T and B cells, to different stimuli, across the different media. Discussion The serum-free media formulations tested in this study offer defined systems for. measuring B cell IgG responses, in vitro, in either a ‘T cell-independent’ (CpG + IL-15) or “T cell-dependent” (PWM or CS) manner and for assessing B cell proliferation, particularly in response to a “T cell-independent” stimulus. However, there are some characteristics and features endowed by human serum that appear to be missing. Therefore, further work is required to optimise animal-free, chemically defined culture conditions for PBMC based assays for inclusion in tiered safety assessments.
View
View
Study on the feasibility of using livestock blood as a fetal bovine serum substitute for cultured meat
Article
Full-text available
Fetal bovine serum (FBS) accounts for the largest portion of the cost of cultured meat production or cell culture experiments and is highly controversial in terms of animal welfare because it is taken from the fetus of a pregnant cow during slaughtering. Nevertheless, FBS is the most important supplement in the cell culture manufacturing process. This study aimed to develop an FBS substitute from slaughterhouse waste blood to reduce the cost of FBS in cultured meat production through various experiments. Our study successfully demonstrated that adult livestock blood obtained from slaughterhouses can effectively replace FBS. Our substitute, when cultured with bovine myosatellite cells, demonstrated cell growth that was either equivalent to or superior to that of commercial FBS. In the process of muscle generation through differentiation, the substitutes from bovine and chicken formed 70%–75% more bovine muscle compared to the control group using FBS. Furthermore, using the FBS substitute can reduce cell culture costs by approximately 61% compared to using commercial FBS. Therefore, the groundbreaking FBS substitute will not only contribute to the development of technology to mass‐produce cultured meat using livestock byproducts but will also lower the production cost of media for experimental cell culture or vaccine production.
View
Analysis of Novel Food Proteins from Plants
Chapter
  • Oct 2024
In the past decades, significant advancement has been made in plant-based platforms for vaccine and pharmaceutical production. Various plant species, including tobacco, corn, potato, and others, have been engineered to produce a wide range of bioactives such as vaccines. Nowadays, with global changes challenging food security for humans, attention of researchers has moved to the use of plant as biotechnological systems for production of nutritious proteins for human nutrition, including animal proteins, that have higher biological values than those from other sources. Although plant-based platforms have considerable advantages over traditional systems such as bacterial and animal systems, there are several obstacles to commercial-scale production, first of all the purification process of the proteins from the expression systems and their structural characterization to ensure their safety and regulatory approval. In this chapter, the issues connected to the production of proteins in plant seeds and a pipeline for their isolation and analysis for proteomics characterization are detailed.
View
Patient-Derived Stem Cell Medicine: A Safe and Reliable Cell Processing and Regenerative Therapeutic Application of Human Dental Pulp Stem Cells
Chapter
  • Sep 2024
Dental tissues obtained from teeth extracted for orthodontic purposes are a safe and easily accessible source of human mesenchymal stem/stromal cells (MSCs), which could be used for future tissue regeneration therapy. Dental tissues are the source of several MSCs, among which dental pulp-derived MSCs (DPSCs) have been studied the most. These cells possess excellent MSC characteristics, such as a high proliferation rate, multipotency, and immunomodulatory properties, and have a very low risk of tumorigenicity. As a result, DPSCs are considered a promising MSC alternative compared to other commonly used MSC sources, such as bone marrow, adipose tissue, and umbilical cords. Tooth extraction for DPSC collection is usually performed in the clinic, even in the absence of immediate regenerative therapeutic requirements. To prepare for possible future clinical use, it is necessary to implement dental cell banking platforms to store these dental MSCs. This book chapter summarizes clinical trials and preclinical animal studies demonstrating recent advances in DPSC-based therapies as treatment approaches for several systemic diseases. These recent advances indicate that clinical-grade cell processing and banking will be necessary to establish future MSC-preservation platforms and to promote safe and efficient clinical applications of patient-derived stem cell medicine.
View
Chemisch definiert – ein zellbasierter Zytotoxizitätsassay ohne fötales Kälberserum
Article
Full-text available
  • Feb 2017
Tests for cytotoxicity–respectively biocompatibility–are performed e. g. for biological evaluation of medical devices or during the development of microphysiometric systems. Here, often cell culture methods which involve the use of fetal bovine serum (FBS) are employed. However, using FBS raises scientific and ethical concerns. In the presented work a protocol for a chemically defined test for cytotoxicity is described.
View
Human Platelet Lysates as a Serum Substitute in Renal Epithelial Cell Culture
Article
  • Apr 2012
Cultured renal epithelia form monolayers of differentiated, polarized cells. On permeable supports, an apical and a basolateral compartment are separated by the cultured epithelium to study epithelial transport in vitro. However, culture conditions and culture media have a significant impact. In this respect, the quality of fetal bovine serum (FBS) seemed to be of major importance. The high lot‐to‐lot variablity of FBS was found to substantially influence the differentiation of epithelial cultures with regard to the generation of a transepithelial electrical resistance (TEER). Thus, batch‐testing of FBS was required. We recently reported on the use of human platelet lysates (PL) as a replacement for FBS. The growth promoting capacity of PL was tested on renal epithelial cell lines, for which differentiation end points are well established. PL support growth, proliferation and differentiation of LLC‐PK1, HK‐ 2 and MDCK cells. In addition, TEER was monitored in filter‐grown LLC‐PK1 and in MDCK II and MDCK I epithelia. Low‐resistance epithelia generated a TEER of 150–250 Ω•cm2 in PL‐media, as seen with FBS, and high‐resistance MDCK I retained their TEER of 8,000 Ω•cm2. PL are a valuable, animal‐derived component‐free substitute for FBS in renal epithelial cell culture with a proven high batch‐to batch uniformity.
View
View
View
Eye Banking in the 21st Century: How Far Have We Come? Are We Prepared for What’s Ahead?
Article
  • Sep 2012
PURPOSE: This review of milestones in eye banking describes efforts by the eye banking community to ensure the quality and quantity of corneal tissue over the last century and anticipates key challenges going forward. METHODS: This account draws on a review of the scientific literature, public documents, and eye banking statistics and is augmented by the recollection of the author, discussions with eye bankers, and an analysis of pressing medical and nonmedical issues that will bear on the future success of eye banking in the United States and internationally. RESULTS: The author identifies five eras of eye banking, highlighting scientific breakthroughs in surgical techniques, storage, and instrumentation; the professionalization of eye banks through development of medical standards and accreditation programs; and the advent of laws and regulations leading to reimbursement changes and donor legislation. The author next identifies five strengths exhibited by the community to achieve those milestones and delineates crucial questions posed by an ever-expanding array of unprecedented demographic, socioeconomic, geopolitical, biological, regulatory, technological, cultural, and ethical challenges. CONCLUSIONS: Technological advances and collaborative efforts have brought the eye banking community to the enviable position it enjoys today. Only by building on its historical strengths can eye banking professionals worldwide successfully evolve their role in an increasingly complex future.
View
View
A consensus introduction to serum replacements and serum-free media for cellular therapies
Article
  • Dec 2016
  • CYTOTHERAPY
The cell therapy industry is a fast-growing industry targeted toward a myriad of clinical indications. As the cell therapy industry matures and clinical trials hit their pivotal Phase 3 studies, there will be a significant need for scale-up, process validation, and critical raw material quality assurance. Part of the well discussed challenges of upscaling manufacturing processes there is a less discussed issue relating to the availability of raw materials in the needed quality and quantities. The FDA recently noted that over 80% of the 66 investigational new drug (IND) applications for mesenchymal stem cell (MSC) products analyzed described the use of FBS during manufacturing. Accumulated data from the past years show an acceleration in serum consumption by at least 10%-15% annually, which suggests that the global demand for serum may soon exceed the supply. Ongoing concerns of safety issues due to risks of various pathogen contaminations, as well as issues related to the aforementioned serum variability that can affect final product reproducibility, are strong motivators to search for serum substitutes or serum-free media. it is important to note that there are no accepted definitions for most of these terms which leads to misleading's and misunderstandings, where the same term might be defined differently by different vendors, manufacturer, and users. It is the drug developer's responsibility to clarify what the supplied labels mean and to identify the correct questions and audits to ensure quality. The paper reviews the available serum replacements, main components, basic strategies for replacement of serum and suggests definitions.
View
Influence of Donor Age and Stimulation Intensity on Osteogenic Differentiation of Rat Mesenchymal Stromal Cells in Response to Focused Low-Intensity Pulsed Ultrasound
Article
  • Sep 2016
A focused low-intensity pulsed ultrasound (FLIPUS) was used to investigate the effects of stimulation period, acoustic intensity and donor age on the osteogenic differentiation potential of rat mesenchymal stromal cells (rMSCs). rMSCs from 3- and 12-mo-old female Sprague Drawly rats were isolated from bone marrow and stimulated 20 min/d with either 11.7 or 44.5 mW/cm(2) (spatial average temporal average intensity) for 7 or 14 d. Osteogenic differentiation markers, i.e., Runt-related transcription factor 2 (RUNX2), osteocalcin (OCN) and degree of matrix calcification were analyzed. On day 7 of stimulation, OCN gene expression was enhanced 1.9-fold in cells from young rats when stimulated with low intensity. The low intensity also led to a 40% decrease in RUNX2 expression on day 7 in aged cells, whereas high intensity enhanced expression of RUNX2 on day 14. FLIPUS treatment with low intensity resulted in a 15% increase in extracellular matrix mineralization in young but not old rMSCs. These differences suggest the necessity of a donor-age related optimization of stimulation parameters.
View
View
Reproducibility: Respect your cells!
Article
  • Sep 2016
p>Numerous variables can torpedo attempts to replicate cell experiments, from the batch of serum to the shape of growth plates. But there are ways to ensure reliability.</p
View