Large-scale expansion of human umbilical cord mesenchymal stem cells in human platelet lysate as a substitute of fetal bovine serum / 中国组织工程研究

ABSTRACT

BACKGROUND:Classic media and fetal bovine serum are commonly used in the culture of umbilical cord mesenchymal stem cells, but the potential risk of serum culture limits its clinical application. OBJECTIVE:To use human platelet lysate alternative to fetal bovine serum for large-scale production of mesenchymal stem cells for therapeutic applications. METHODS:Human platelet lysate was prepared by repeated freezing and thawing, centrifugation, filtration, and concentration. Human umbilical cord mesenchymal stem cells were cultured in Iscove’s Modified Dulbecco’s Medium containing 5%concentrated platelet lysate (experimental group) or 10%fetal bovine serum (control group). After separation and enzymatic digestion, human umbilical cord mesenchymal stem cells were subcultured at a concentration of 3 000/cm2 up to the fifth generation. Then, cellmorphology and diameter, immune phenotype, osteogenic and adipogenic differentiation, and cloning efficiency were detected and compared between two groups. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a smal er size and more elongated morphology than those in fetal bovine serum-supplemented media. Colony forming unit-fibroblast analyses further showed no significant differences in colony efficiency. Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed an increase of proliferation capacity;whereas, similar immunophenotypes remained in the two groups. In vitro assays revealed intact differentiation potential. Moreover, human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a significantly higher capacity to differentiate towards osteocytes, indicating human platelet lysate is an alternative to fetal bovine serum for low-density production of mesenchymal stem cells for therapeutic applications.