山东医药2018，Vol.58Issue(12)：9-13,5.DOI:10.3969/j.issn.1002-266X.2018.12.003
BMP-2对间充质干细胞促迸造血干细胞增殖的影响及其机制
Synergistic effect of bone marrow mesenchymal stem cell and BMP-2 on proliferation of hematopoietic stem cells
摘要
Abstract
Objective To investigate the role of human bone morphogenetic protein 2(BMP-2)in mesenchymal stem cells(MSCs)promoting the proliferation of hematopoietic stem cells(HSCs)and its possible mechanism.Methods The whole bone marrow adherent culture method was used to expand MSC from primary to the third passage,MSCs were identi-fied by flow cytometry with a purity of 52.4%;the human bone marrow CD34 +cells(that is HSC)were sorted by mini MACS magnetic bead separator from adults′bone marrow,and the purity of the HSCs was 81.9%identified by flow cytom-etry.The HSCs were non-contact co-cultured with the third passage of MSCs by Transwell.On day 3,7,and 10,we coun-ted the number of HSCs.The fastest proliferation time of HSCs was the seventh day of co-culture.In the next experiment, the cells were divided into six groups: the HSC group and MSC group(we used the single cell culture), HSC+BMP-2 group and MSC+BMP-2 group(which were added with 100 ng/mL BMP-2),HSC+MSC group(the two groups of cells were co-cultured), and the HSC +MSC+BMP-2 group(HSCs were co-cultured with MSCs and then were added with 100 ng/mL BMP-2).On day 7 of culture,the number of HSCs was counted by using cell counting plate,and the prolifera-tion index Ki67 and the expression of angiogenesis-related factors Tie2,VEGF,and Ang1 mRNA was detected by real-time fluorescent quantitative qRT-PCR.Results The number of HSCs in the HSC group, HSC+BMP-2 group, HSC+MSC group,HSC+MSC+BMP-2 group was respectively(15.37 ±0.42)×104,(37.81 ±1.07)×104,(117.00 ±2.53)× 104, and(129.06 ±5.13)×104,and significant difference was found between groups(P<0.05 or P<0.01).The rela-tive expression of Ki67 and Tie2 mRNA of HSCs in the HSC group,HSC+BMP-2 group,HSC+MSC group,and HSC+MSC+BMP-2 group increased gradually.Similarly, the VEGF and Ang1 mRNA relative expression of MSCs in the MSC group,MSC+BMP-2 group, MSC +HSC group, and MSC +HSC +BMP-2 group increased gradually, and significant difference was found between groups(P<0.05 or P<0.01).Conclusion BMP-2 could promote the proliferation effect of MSC on HSC by enhancing the expression of angiogenesis-related factors VEGF,Ang1,and Tie2.关键词
造血干细胞/间充质干细胞/人骨形态发生蛋白2/细胞增殖Key words
hematopoietic stem cells/mesenchymal stem cells/human bone morphogenetic protein 2/cell prolifera-tion分类
医药卫生引用本文复制引用
李书坛,黄远琼,汪姝玥,林凡莉,黄纯兰..BMP-2对间充质干细胞促迸造血干细胞增殖的影响及其机制[J].山东医药,2018,58(12):9-13,5.基金项目
国家自然科学基金资助项目(81450030) （81450030）
四川省教育厅重大培育项目(14CZ0017). （14CZ0017）
