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

被引:39
作者
Cowper, Mathew [1 ,7 ]
Frazier, Trivia [1 ,2 ,3 ]
Wu, Xiying [2 ,3 ]
Curley, J. Lowry [2 ,4 ]
Ma, Michelle H. [3 ]
Mohiuddin, Omair A. [1 ]
Dietrich, Marilyn [5 ]
McCarthy, Michelle [1 ]
Bukowska, Joanna [1 ,6 ]
Gimble, Jeffrey M. [1 ,2 ,3 ]
机构
[1] Tulane Univ, Sch Med, New Orleans, LA 70112 USA
[2] LaCell LLC, New Orleans, LA 70148 USA
[3] Obatala Sci Inc, New Orleans, LA 70148 USA
[4] Axosim Inc, New Orleans, LA 70112 USA
[5] Louisiana State Univ, Sch Vet Med, Baton Rouge, LA 70803 USA
[6] Polish Acad Sci, Inst Anim Reprod & Food Res, PL-10748 Olsztyn, Poland
[7] Wake Forest Univ, Bowman Gray Sch Med, Dept Urol, Winston Salem, NC 27101 USA
关键词
adipogenesis; adipose-derived stromal/stem cells; chondrogenesis; colony forming unit-fibroblast; fetal bovine serum; human platelet lysate; mesenchymal stem cell; osteogenesis; regenerative medicine; MESENCHYMAL STEM-CELLS; VASCULAR FRACTION; COLONY FORMATION; EXPANSION; DIFFERENTIATION; PROLIFERATION; SECRETOME;
D O I
10.3390/cells8070724
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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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页数:13
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