内皮祖细胞条件培养基对间充质干细胞增殖和成骨分化的影响及其机制

doi:10.13481/j.1671-587x.20150203

Abstract

Abstract:

Objective To investigate the effects of bone marrow-derived endothelial progenitor cells(EPCs) conditioned medium(EPCs-CM) on the proliferation and osteogenic differentiation of mesenchymal stem cells(MSCs),and to clarify the mechanisms.Methods The EPCs and MSCs were isolated from bone marrow of the mice using differential adhesion method.The surface markers of EPCs and MSCs were identified by flow cytometry (FCM).The osteogenic,chondrogenic,and adipogenic induction differentiation abilities of the MSCs were identified.The function of EPCs was identified by tube formation experiment.The MSCs were divided into 0% EPCs-CM group(control group,cultured with LG-DMEM),50% EPCs-CM group(cultured with 50% EPGs-CM and 50% LG-DMEM),and 100% EPCs-CM group(cultured with 100% EPCs-CM).The proliferation activities of the MSCs in various groups were detected by MTT method;the osteogenic differentiation abilities of the MSCs in various groups were detected by Alizarin red staining.Results The FCM results showed that the third passage MSCs were strongly positive for Sca-1,CD29 and negative for CD45,CD11b and could be induced to complete differentiation process into osteoblasts and adipocytes and chondrocytes.The third passage EPCs cultured on Matrigel showed tube-like structures and highly expressed CD34,CD133 and VEGFR2.Compared with control group,the proliferation activities of the MSCs in 50% EPCs-CM group and 100% EPCs-CM group were increased(P<0.05),which presented a dose-dependent manner.The Alizarin red staining results showed the number of mineralized nodules and calcium deposition of the MSCs in 50% EPCs-CM group and 100% EPCs-CM group were higher than those in control group after cultured for 21 d(P<0.05).Conclusion The method of differential adhesion can simultaneously isolate the MSCs and EPCs.EPCs-CM can promote the proliferation and osteogenic differentiation of the MSCs.

Key words: mesenchymal stem cells, endothelial progenitor cells, conditioned medium, cell proliferation, osteogenic differentiation

CLC Number:

R683

FENG Wenlei, ZHANG Meng, XU Fangjie, YIN Shuanghong, WANG Yanjie, CHEN Xueling, WU Xiangwei. Effects of endothelial progenitor cells conditioned medium on proliferation and osteogenic differentiation of mesenchymal stem cells and their mechanisms[J].Journal of Jilin University Medicine Edition, 2015, 41(02): 218-224.

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Effects of endothelial progenitor cells conditioned medium on proliferation and osteogenic differentiation of mesenchymal stem cells and their mechanisms

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