定向诱导骨髓间充质干细胞向心肌细胞分化

Induction and differentiation of rat marrow-derived mesenchymal stem cells into cadiocytes in vitro

目的:采用骨髓间充质干细胞体外转化为心肌样细胞,为心衰的干细胞移植治疗提供基础。方法:分离培养大鼠骨髓间充质干细胞,5-氮杂胞苷诱导24h后正常培养,通过形态学、PAS反应、磷钨酸苏木素染色(PTAH)、免疫细胞化学染色作鉴定。结果:诱导细胞以梭形、柱状为主,部分细胞有分支,单个核、卵圆形、居中,似心肌细胞,PAS及PTAH染色均为阳性。诱导后培养2周的细胞表达Sarcomeric Actin和Connexin-43较弱,以后表达逐渐增强,而且1、3、5代细胞均稳定表达。结论:骨髓MSCs可在体外被诱导分化为心肌样细胞,有望成为心衰干细胞移植治疗的理想细胞材料。

Objective： To differentiate rat marrow-derived mesenchymal stem cells （MSCs） to cadiocytes in vitro via the induction of 5-azacytidine and provide foundation for serving MSCs as a new stem cell source to treat heart failure. Methods： Rat MSCs were isolated and cultured. After induced with 5-azacytidine for 24 hours and then normal cultured, cells were identified by morphology, periodic-acid-schiff reaction （PAS）, phosphotungstic acid hematoxylin （PTAH） and immunocytochemistry. Results： Most MSCs induced gradually became spindle-like and pillar-form with oval nuclei lying in the center of cells and similar to cadiocytes, and being of positive reaction in PAS and PTAH. In addition, more than 2 weeks of cultivation, positive staining of Sarcomeric Actin and Connexin-43 were observed on those cells, and the same expressions were on P1, P3 and P5 cells, Conclusions： MSCs can be induced into cadiocytes by 5-azacytidine in vitro, which suggests that it might be ideal donor cells in stem cell transplantation for heart failure therapy.