不同来源血清体外培养骨髓间充质干细胞的研究

Study on amplification of bone marrow mesenchymal stem cells with serum from different sources in vitro

目的适当的血清浓度可维持骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）的体外培养，而不同来源的血清对BMSCs的培养效果是否不同。本研究探讨自体血清、脐带血清、AB血清、胎牛血清在体外培养人BMSCs的可行性。方法Ficoll密度梯度离心法结合贴壁法分离纯化人BMSCs，在培养过程中分别加入自体血清、脐带血清、AB血清、胎牛血清，观察比较细胞形态、表面抗原、生长曲线及各组在诱导剂作用下表面抗原和诱导分化的相关性。结果从BMSCs的形态学、生长曲线、表面抗原、诱导分化上看各组无明显差异，生长状况良好。结论自体血清、脐带血清、AB血清、胎牛血清均可维持BMSCs的体外培养，而白体血清解决异种和异体血清带来的一些潜在的风险，更为安全。

Objective To investigate whether autologous serum could replace fetal calf serum for the expansion of human human bone marrow mesenchymal stem cells （BMSCs）in vitro and establish a protocol of culturing BMSCs. Methods BMSCs were separated from human bone marrow by modified ficoll density gradient centrifugation combined with adherence culture method and respectively cultured by DMEM/F12 with 10% umbilical cord blood serum, 10% fetal calf serum, human AB blood serum, and autologous serum in vitro. The morphology, and its growth character of the BMSCs were studied with the phase contrast microscope. Differentiation potency through condition medium directional induction were observed. The expression of cell surface antigens was detected by flow cytometer, immunocytochemistry. Results There were no significant differences between four groups in morphological changes. BMSCs were induced to differentiate into neuron like cells by growth factor. The induced cells expressed NSE, NF, GFAP after induction. Conclusion High purity and active BMSCs are cultured by autoserum, which grow prosperously and maintain its biological characters for at least 5 passages.