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

• Main Authors: Mathew Cowper, Trivia Frazier, Xiying Wu, J. Lowry Curley, Michelle H. Ma, Omair A. Mohiuddin, Marilyn Dietrich, Michelle McCarthy, Joanna Bukowska, Jeffrey M. Gimble
• Format: Article
• Language: English
• Published: MDPI AG 2019-07-01
• Series: Cells
• Subjects: adipogenesis; adipose-derived stromal/stem cells; chondrogenesis; colony forming unit-fibroblast; fetal bovine serum; human platelet lysate; mesenchymal stem cell; osteogenesis; regenerative medicine
• Online Access: https://www.mdpi.com/2073-4409/8/7/724

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.