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

Cells. 2019 Jul 15;8(7):724. doi: 10.3390/cells8070724.

Abstract

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.

Keywords: adipogenesis; adipose-derived stromal/stem cells; chondrogenesis; colony forming unit-fibroblast; fetal bovine serum; human platelet lysate; mesenchymal stem cell; osteogenesis; regenerative medicine.

MeSH terms

  • Adipose Tissue / cytology*
  • Animals
  • Blood Platelets / chemistry*
  • Cattle
  • Cell Differentiation*
  • Cell Proliferation
  • Cells, Cultured
  • Culture Media, Serum-Free / chemistry*
  • Culture Media, Serum-Free / pharmacology
  • Humans
  • Mesenchymal Stem Cells / cytology*
  • Mesenchymal Stem Cells / drug effects
  • Mesenchymal Stem Cells / physiology
  • Serum / chemistry

Substances

  • Culture Media, Serum-Free