title: In vitro proliferation and differentiation of human mesenchymal stem cells cultured in autologous plasma derived from bone marrow.
abstract: Expansion of human mesenchymal stem cells (hMSCs) in medium supplemented with fetal bovine serum (FBS) has a potential risk of transmitting viral and prion diseases and causing immunological rejection. The aim of our present study was to find a substitute for the traditional FBS in culture of hMSCs to facilitate the clinical application of hMSCs. We used autologous plasma derived from bone marrow (APM) as a substitute for FBS and found that, when cultured with APM, the cell surface markers and the proportion of hMSCs in the G(0)/G(1) phase and the S+G(2)/M phase resembled those cultured with FBS. However, there were fewer early apoptotic cells in APM-supplemented medium than in FBS (p < 0.01). APM resulted in much greater thymidine incorporation than FBS (p < 0.001). There were significantly more alkaline phosphatase (ALP)-positive fibroblast colony-forming units (CFU-Fs) covering larger areas in APM than in FBS (p < 0.01). Also, APM induced greater ALP activity, more mineralized nodules, and greater expression of osteogenic genes than did FBS. In addition, when cultured in adipogenic medium, there were fewer oil-red O-positive cells and lower expression of adipogenic gene with APM than with FBS. In conclusion, expansion of hMSCs in APM-supplemented medium instead of traditional FBS is more advantageous. It could promote cell proliferation, enhance osteogenic differentiation, and suppress adipogenic differentiation of hMSCs and is therefore a safer and more effective substitute for FBS in clinical cytotherapy processes.
authors.affiliation: Department of Orthopedic Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, PR China.
authors.forename: Xiaojiang
authors.initials: X
authors.lastname: Sun
authors.forename: Yaokai
authors.initials: Y
authors.lastname: Gan
authors.forename: Tingting
authors.initials: T
authors.lastname: Tang
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Zhang
authors.forename: Kerong
authors.initials: K
authors.lastname: Dai
mesh_terms: D050156:Adipogenesis / Q000235:genetics; D000328:Adult; D000469:Alkaline Phosphatase / Q000378:metabolism; D017209:Apoptosis; D001853:Bone Marrow / Q000502:physiology*; D002452:Cell Count; D018929:Cell Culture Techniques; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000201:enzymology; D008875:Middle Aged; D010012:Osteogenesis / Q000235:genetics; D010949:Plasma / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D044967:Serum; D013194:Staining and Labeling; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
chemical_list: D014316:Tritium; D000469:Alkaline Phosphatase; D013936:Thymidine