Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells

Introduction: Adipose derived stromal/stem cells (ASCs) hold potential as cell therapeutics for a wide range of disease states; however, many expansion protocols rely on the use of fetal bovine serum (FBS) as a cell culture nutrient supplement. The current study explores the substitution of lysates from expired human platelets (HPLs) as an FBS substitute. Methods: Expired human platelets from an authorized blood center were lysed by freeze/thawing and used to examine human ASCs with respect to proliferation using hematocytometer cell counts, colony forming unit-fibroblast (CFU-F) frequency, surface immunophenotype by flow cytometry, and tri-lineage (adipocyte, chondrocyte, osteoblast) differentiation potential by histochemical staining. Results: The proliferation assays demonstrated that HPLs supported ASC proliferation in a concentration dependent manner, reaching levels that exceeded that observed in the presence of 10% FBS. The concentration of 0.75% HPLs was equivalent to 10% FBS when utilized in cell culture media with respect to proliferation, immunophenotype, and CFU-F frequency. When added to osteogenic, adipogenic, and chondrogenic differentiation media, both supplements showed appropriate differentiation by staining. Conclusion: HPLs is an effective substitute for FBS in the culture, expansion and differentiation of human ASCs suitable for pre-clinical studies; however, additional assays and analyses will be necessary to validate HPLs for clinical applications and regulatory approval.

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Stimulatory Role of Glycated Fetal Bovine Serum Along with Iron on In Vitro Production of Insulin by Differentiated Mouse Embryonic Stem Cells

JOURNAL OF HEALTH SCIENCE ◽

10.1248/jhs.57.356 ◽

2011 ◽

Vol 57 (4) ◽

pp. 356-361

Author(s):

Ikuo Nishigaki ◽

Gowri Rangasamy Gunassekaran ◽

Panjan Nagappan Venkatesan ◽

Mandupal Chaco Sabu ◽

Sabu Priya ◽

...

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Embryonic Stem ◽

In Vitro Production ◽

Fetal Bovine ◽

Vitro Production

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241 9.22 Performance of Gamma-irradiated Fetal Bovine Serum in Cell Culture

Animal Cell Technology ◽

10.1016/b978-0-7506-1845-8.50023-5 ◽

1994 ◽

pp. 99-101

Author(s):

J.D. Keathley ◽

D. Wyatt ◽

C.M. Williams ◽

R. Festen ◽

C. Maben

Keyword(s):

Cell Culture ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Fetal Bovine ◽

Gamma Irradiated

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Elucidation of the Interaction between Flavan-3-ols and Bovine Serum Albumin and Its Effect on Their In-Vitro Cytotoxicity

Molecules ◽

10.3390/molecules24203667 ◽

2019 ◽

Vol 24 (20) ◽

pp. 3667

Author(s):

Yasuyuki Fujii ◽

Yoshitomo Suhara ◽

Yusuke Sukikara ◽

Tomohiro Teshima ◽

Yoshihisa Hirota ◽

...

Keyword(s):

Cell Culture ◽

Bovine Serum Albumin ◽

Serum Albumin ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

In Vitro Cytotoxicity ◽

Binding Energies ◽

Docking Simulations ◽

Fetal Bovine

Flavan-3-ols (FLs), specifically catechin and its oligomer B-type procyanidins, are suggested to potently bind to bovine serum albumin (BSA). We examined the interaction between BSA and FLs by fluorescence quenching and found the following order of binding activities to BSA: cinnamtannin A2 (A2; tetramer) > procyanidin C1 (C1; trimer) ≈ procyanidin B2 (B2, dimer) > (−)epicatechin (EC, monomer). Docking simulations between BSA and each compound at the binding site showed that the calculated binding energies were consistent with the results of our experimental assay. FLs exerted cytotoxicity at 1000 μg/mL in F11 cell culture with fetal bovine serum containing BSA. In culture containing serum-free medium, FLs exhibited significant cell proliferation at 10−4 μg/mL and cytotoxicity was observed at concentrations greater than 10 μg/mL. Results of this study suggest that interactions between polyphenols and BSA should be taken into account when evaluating procyanidin in an in vitro cell culture system.

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Detection of bovine viruses in fetal bovine serum used in cell culture

In Vitro ◽

10.1007/bf02616377 ◽

1975 ◽

Vol 11 (6) ◽

pp. 400-403 ◽

Cited By ~ 34

Author(s):

A. J. Kniazeff ◽

L. J. Wopschall ◽

H. E. Hopps ◽

C. S. Morris

Keyword(s):

Cell Culture ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Fetal Bovine ◽

Bovine Viruses

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302 REPLACEMENT OF FETAL BOVINE SERUM WITH KNOCKOUT SERUM REPLACER AND STEM CELLS CONDITIONED MEDIUM DURING IN VITRO CULTURE OF BOVINE EMBRYOS

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab302 ◽

2010 ◽

Vol 22 (1) ◽

pp. 307

Author(s):

M. M. Souza ◽

N. Z. Saraiva ◽

C. S. Oliveira ◽

T. A. D. Tetzner-Nanzeri ◽

R. Vantini ◽

...

Keyword(s):

Stem Cells ◽

In Vitro Culture ◽

Bovine Serum ◽

Fetal Bovine Serum ◽

Protein Supplementation ◽

Control Group ◽

Bovine Embryos ◽

Fetal Bovine

The use of fetal bovine serum (FBS) as protein supplementation in IVP of bovine embryos has presented difficulties because it can introduce a number of pathogenic components in culture systems, can be related to the birth of calf with abnormal growth and development, and precludes the establishment of the actual nutritional needs of the embryo, because it contains an unlimited variety of substances. This study evaluated the replacement of the FBS in the medium of in vitro culture (IVC) of bovine embryos, using the knockout serum replacer (KSR) as protein supplementation and culture medium conditioned with stem cells. Therefore, bovine oocytes from ovaries of slaughterhouse were selected and matured in vitro in TCM-199 medium supplemented with 10% FBS (Crypion), 1.0 μg mL-1 FSH (Pluset®, Calier, Barcelona, Spain), 50 μg mL-1 hCG (Profasi®, Serono, Geneva, Switzerland), 1.0 μg mL-1 estradiol (Sigma E-2758, Sigma Chemical, St. Louis, MO, USA), 0.2 mM sodium pyruvate, and 83.4 μg mL-1 amikacin for 24 h. After that, 1144 oocytes were fertilized in IVF-TALP medium containing 6 mg mL-1 of BSA. After 18 to 22 h, the zygotes were cultured in SOF + 5% FBS (group 2); SOF + 5% KSR (group 3); SOF (5% FBS) + 10% SOF (5% FBS) conditioned by stem cells (group 4); or SOF (5% KSR) + 10% SOF (5% KSR) conditioned by stem cells (group 5), in an atmosphere of 5% O2 at 38.5°C for 8 days. A control group outside the controlled atmosphere was added, supplemented with 5% FBS (group 1). The SOF medium supplemented with 5% FBS or KSR was conditioned by stem cells and added to SOF medium for the culture of embryo at a concentration of 10%. The rates of cleavage and production of blastocysts were assessed 48 hours and 7 days after IVF, respectively, and analyzed by chi-square test, with a significance level of 5% in the statistical program Minitab® (release 14.1, Minitab, State College, PA, USA). On the eighth day, the TUNEL test for determination of the percentage of apoptosis and the differential staining technique for determination of inner cell mass (ICM) and trophoblast (TF) were performed. The results were submitted to ANOVA, followed by comparing the means by Tukey’s test using the program GraphPad Prism (GraphPad, San Diego, CA, USA). The treatments did not differ in the production of embryos, being similar to the control group: G1 = 31.75% (74/233), G2 = 35.26% (79/224), G3 = 32.70% (74/226), G4 = 28.76% (63/219), and G5 = 26.85% (65/242). With regard to the assessment of embryonic quality, the treatments showed similar results to the control groups. No differences were observed among groups both in color and ICM/TF ratio (G1 = 0.60, G2 = 0.62, G3 =0.65, G4 = 0.60, and G5 = 0.60). Furthermore, the TUNEL showed no significant difference in the percentage of apoptosis among groups (G1 = 7.10%, G2 = 3.76%, G3 = 5.58%, G4 = 4.50%, and G5 = 4.11%). The data obtained so far indicate that it is possible to produce embryos in vitro by replacing the FBS in the culture, achieving results similar to those obtained with serum. Financial support: FAPESP 2007/58506-6.

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