大鼠骨髓源性间充质干细胞对内皮细胞增殖及成管的影响

The effect of rat bone marrow-derived mesenchymal stem cells on proliferation and angiogenesis of endothelial cells

目的观察大鼠骨髓源性间充质干细胞（MSCs）对内皮细胞（ECs）增殖及成管能力的影响。方法采用密度梯度离心、流式细胞仪及免疫荧光的方法将不同培养条件下（DMEM及EGM-2培养液）大鼠骨髓源性单核细胞（BM—MNCs）进行分离及鉴定，并对直接和间接共培养（Transwells）的细胞进行体外成管和免疫荧光统计分析。结果与DMEM中的细胞比较，EGM-2培养的BM—MNCs能够形成典型的集落形成细胞。流式细胞仪检测和免疫荧光鉴定表明培养在DMEM和EGM-2中的细胞分别为MSCs（CD105、CD90和CD73表达率大于95％）和ECs（CD31阳性率大于90％）。与间接共培养比较，直接共培养后细胞的成管数量及长度均显著降低（P〈0．05）。免疫荧光染色进一步证明，直接共培养后ECs的数量显著减少（P〈0．05）。结论直接共培养下，大鼠MSCs能够抑制ECs的增殖及成管。

Objective To investigate the effect of rat bone marrow-derived mesenchymal stem cells （MSCs） on proliferation and angiogenesis of endothelial cells （ECs）. Methods The rat bone marrow-derived mononuclear cells （BM-MNCs） were collected by density gradient eentrifugation and cultured in different medium （DMEM and EGM-2） before identified by flow cytometry or immunofluorescence. After direct and indirect co-culture, they were analysised by vascular network formation in vitro and immunofluorcscence. Results Compared with DMEM, the BM-MNCs could formed typical colony-forming cells in EGM-2. After cultured in DMEM and EGM-2 respectively, MNCs could develop into MSCs with greater than 95% of cells expressing CD105, CD90 and CD73, and ECs with greater than 90% of cells positive staining for CD31. Compared with indirect co-culture, direct co-culture of MSCs and ECs cells would result in significant reduction of tube quantity and tube length （P 〈 0. 05 ）. Furthermore, immunofluorescence staining demonstrated the number of ECs also significantly reduced （ P 〈 0. 05 ）. Conclusion Directly co- cultured MSCs were able to inhibit ECs proliferation and vascular network formation.