细胞间直接接触对骨髓间充质干细胞分化为心肌细胞作用的研究

Study of the effect of cell-cell direct contact to the bone marrow stromal cells differentiate into cardiomyocytes

目的探讨体外心肌细胞(cardiomyocytes,CM)与骨髓间充质干细胞(bone marrow stromal cells,MSCs)共培养时,骨髓间充质干细胞与心肌细胞间直接接触对其分化为心肌细胞的影响。方法API(4,6-二咪基-2-苯吲哚盐酸)标记后,分别与心肌细胞按接触组和非接触(millicellTM-HA半透膜)组培养,1周后应用免疫荧光检测MSCs肌钙蛋白T(cTnT)的表达,并用RT-PCR方法检测NKx-2.5、GATA-4、TGF-β的表达。结果接触培养组中可见DAPI标记的骨髓间充质干细胞表达cTnT,NKx-2.5、GATA-4、TGF-β有高表达,非接触组的MSCs未见cTnT的表达,相关基因仅有低表达。结论心肌细胞与MSCs的直接接触,是诱导MSCs分化为心肌细胞的重要因素。

Objective To investigate the effect of cell-cell direct contact to the bone marrow stromall cells （MSCs） differentiate into cardiomyocytes（CM） when MSCs co-cultured with cardiomyocytes in vitro .Methods The 5-6th generation of MSCs was isolated and purified, after labeled by DAPI, cultured with cardiomyocytes through direct and indirect （millicell^TM-HA semipermeable membrane ）contact. One week later, cell immunofluorescence was carried to detect the expression of cTnT of MSCs,using RT-PCR to investigate the expression of NKx-2.5, GATA-4, TGF-β. Results There was high expression of NKx-2.5, GATA-4, TGF-β of the MSCs in direct contact group, cTnT is positive, and there was low expression of NKx-2.5, GATA-4, TGF-β in indirect contact group, and cTnT expression was negative. Conclusion It demonstrates that the direct contact of MSCs and cardiomyocyte is essential for MSCs differentiation into cardiomyocytes.