两种体外培养体系对人骨髓间充质干细胞原代培养、增殖和成骨分化的影响比较  

Comparison of effect in two vitro culture systems in supporting human bone marrow mesenchymal stem cells for primary culture,propagation and osteogenic differentiation

作　　者：徐杨[1] 林飞跃[1] 尹晓明[1] XU Yang;LIN Feiyue;YIN Xiaoming(Department of Orthopedic Surgery,Fujian Provincial Hospital,Provincial Clinical Medical College of Fujian Medical University,Fuzhou,Fujian 350001,China)

机构地区：[1]福建医科大学省立临床医学院福建省立医院骨一科

出　　处：《福建医药杂志》2019年第6期136-141,F0004,共7页Fujian Medical Journal

基　　金：福建省自然科学基金项目(2016J01505)

摘　　要：目的研究使用混合血小板裂解液(pHPL)和StemPro^■ MSC无血清培养体系对人骨髓间充质干细胞原代培养、增殖和成骨分化的影响。方法分别使用异体浓缩血小板制备的混合血小板裂解液pHPL和StemPro^■ MSC无血清培养体系原代培养人骨髓间充质干细胞,以经典的胎牛血清培养方案做对照,比较3组中的细胞形态、表面标志物、细胞骨架、细胞周期和增殖率,并在加入经典成骨诱导液后检测3组中碱性磷酸酶含量、成骨相关基因含量和矿化结节形成,比较其成骨分化能力。结果pHPL、StemPro^■ MSC和FBS培养组均能较好支持人骨髓间充质干细胞的原代培养,3组中细胞形态、表面标志物、细胞骨架、细胞周期均无明显差异,pHPL和StemPro^■ MSC组中细胞增殖率无明显差异,但均明显较FBS组高。在进行成骨诱导后,pHPL组中碱性磷酸酶含量、成骨相关基因含量均较StemPro^■ MSC组和FBS组高,3组中均能见到成熟的矿化结节形成。结论pHPL和StemPro^■ MSC均能够很好地支持人骨髓间充质干细胞的原代培养,并且促进细胞增殖的能力均较FBS为高,而且pHPL在促进其成骨分化方面较StemPro^■ MSC和FBS更强。Objective To investigate effect of pooled human platelet lysate(pHPL)and StemPro^■ MSC serum-free media in primary culture,propagation and osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMMSCs).Methods pHPL and StemPro^■ MSC were used for primary culture of hBMMSCs,comparing with FBS method.Cell morphology,surface marker expression,cytoskeleton,cell cycle and cell proliferation rate were compared among the three groups.The content of alkaline phosphatase,the content of osteogenic related genes and the formation of mineralized nodule were detected after adding classical osteogenic induction solution,to compare their osteogenic differentiation ability among the three groups.Results Both pHPL and StemPro^■ MSC could support primary culture of hBMMSCs as well as FBS.There were no differences between the three groups in cell morphology,surface marker expression,cytoskeleton and cell cycle.Cell proliferation rate wasn?t significantly higher in pHPL group and StemPro^■ MSC group,but it was significantly higher in both pHPL group and StemPro^■ MSC group than FBS group.The content of alkaline phosphatase and the content of osteogenic related genes were significantly higher in pHPL group than both StemPro^■ MSCgroupand FBS group.The formation of mineralized nodule could be seen in all three groups.Conclusion PHPL and StemPro^■ MSC can well support the primary culture of human bone marrow mesenchymal stem cells,the ability to promote cell proliferation is higher than that of FBS,and pHPL is stronger in promoting osteogenic differentiation than StemPro^■ MSC and FBS.

关 键 词：人血小板裂解液 无血清培养 骨髓间充质干细胞 原代培养 增殖 成骨分化 

分 类 号：R329.2+8[医药卫生—人体解剖和组织胚胎学]