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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...

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Autores principales: Cowper, Mathew, Frazier, Trivia, Wu, Xiying, Curley, J. Lowry, Ma, Michelle H., Mohiuddin, Omair A., Dietrich, Marilyn, McCarthy, Michelle, Bukowska, Joanna, Gimble, Jeffrey M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6679214/
https://www.ncbi.nlm.nih.gov/pubmed/31311198
http://dx.doi.org/10.3390/cells8070724
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author Cowper, Mathew
Frazier, Trivia
Wu, Xiying
Curley, J. Lowry
Ma, Michelle H.
Mohiuddin, Omair A.
Dietrich, Marilyn
McCarthy, Michelle
Bukowska, Joanna
Gimble, Jeffrey M.
author_facet Cowper, Mathew
Frazier, Trivia
Wu, Xiying
Curley, J. Lowry
Ma, Michelle H.
Mohiuddin, Omair A.
Dietrich, Marilyn
McCarthy, Michelle
Bukowska, Joanna
Gimble, Jeffrey M.
author_sort Cowper, Mathew
collection PubMed
description 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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spelling pubmed-66792142019-08-19 Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells Cowper, Mathew Frazier, Trivia Wu, Xiying Curley, J. Lowry Ma, Michelle H. Mohiuddin, Omair A. Dietrich, Marilyn McCarthy, Michelle Bukowska, Joanna Gimble, Jeffrey M. Cells Brief Report 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. MDPI 2019-07-15 /pmc/articles/PMC6679214/ /pubmed/31311198 http://dx.doi.org/10.3390/cells8070724 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Cowper, Mathew
Frazier, Trivia
Wu, Xiying
Curley, J. Lowry
Ma, Michelle H.
Mohiuddin, Omair A.
Dietrich, Marilyn
McCarthy, Michelle
Bukowska, Joanna
Gimble, Jeffrey M.
Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title_full Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title_fullStr Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title_full_unstemmed Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title_short Human Platelet Lysate as a Functional Substitute for Fetal Bovine Serum in the Culture of Human Adipose Derived Stromal/Stem Cells
title_sort human platelet lysate as a functional substitute for fetal bovine serum in the culture of human adipose derived stromal/stem cells
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6679214/
https://www.ncbi.nlm.nih.gov/pubmed/31311198
http://dx.doi.org/10.3390/cells8070724
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