骨髓间充质干细胞对与平滑肌细胞共培养体系中细胞增殖和自身分化的影响

Influence of marrow mesenchymal stem cells on cell proliferation and self differentiation in a direct co-culture system with smooth muscle cell

目的探讨骨髓间充质干细胞(marrow mesenchymal stem cells,MSCs)对联合培养细胞增殖及自身表达平滑肌肌动蛋白(smooth muscle alpha-actin,α-actin SM)的影响。方法建立MSCs与平滑肌细胞(smooth muscle cells,SMCs)直接共培养体系,对MSCs与SMCs直接共培养组和单纯SMCs组及MSCs组在不同时间点细胞计数。对共培养组和单纯MSCs组行4’6’-二乙酰基-2-苯基吲哚(DAPI)(5 ng/ml)标记,利用抗α-actin SM荧光抗体,检测2组MSCs胞浆内-actin SM表达。结果共培养组细胞计数较单独SMCs培养组明显减少,以第4天为著(P<0.01),与SMCs共培养5 d,MSCs在共培养组较单纯MSCs组表达α-actin SM阳性率明显降低(P<0.05)。结论在无干预因素情况下,MSCs与SMCs直接共培养明显抑制共培养体系中细胞增殖,且抑制MSCs表达α-actin SM。

Objective To investigate the influence of marrow mesenchymal stem cells （MSCs） on cell proliferation and self differentiation in a co-culture system with smooth muscle cell （SMCs） and self-expression of smooth muscle alpha-actin （α-actin SM ）. Methods The direct co-cuhure system of MSCs with SMCs was established, and the cell count was performed in co-culture group, simple SMCs group and simple MSCs group at different time points. The cells in co-culture group and MSCs group were pre-labled by DAPI （5ng/ml）, and the expression of ct-actin SM was detected with anti-α-actin SM antibody by fluorescence immunocytochemistry for both groups. Results The cell counts in direct co-culture group were significantly lower than those in simple SMCs group,especially on the 4th day （ 1.69 × 10^4/ml vs 27.20× 10^4/ml, P 〈 0.01 ）, after 5-day co-culture with SMCs,the positive rate of α-actin SM expression of MSCs in co-culture group was obviously decreased,as compared with that in simple MSCs group （44.1% vs 26.6%, P 〈 0.05 ）. Conclusion MSCs in the direct co-culture system with SMCs in vitro can significantly inhibit cell proliferation and reduce the α-actin SM expression of MSCs,if there is no intervention factor.