摘要
目的:建立成肌细胞与心肌细胞体外联合培养的模型,并研究其合胞体的形成。 方法:取1-3日龄新生大鼠,在体外分别分离提纯成肌细胞和心肌细胞,以1:1细胞数混合(浓度均为1×10~8 L^(-1))接种于培养板中,在联合培养的第4天收集细胞做电镜观察,活细胞玻片做微注射活化生物素,以德科萨斯红标记的卵白素作为二抗标记,荧光显微镜观察。结果:成肌细胞有约95％的细胞抗α-肌动蛋白,结蛋白染色阳性,在透射电镜观察成肌细胞与心肌细胞联合培养的细胞超微结构,发现了两种细胞能够形成桥粒连接,未见到间隙连接,而细胞微注射则显示活化生物素可以通过两种细胞形成的连接,即间隙连接。 结论:在体外,两种细胞联合培养可以形成桥粒连接和间隙连接,即电机械连接,为成肌细胞的心脏内移植可能形成合胞体提供了一个研究方法和有力证据。
AIM: To establish a model co-cultured with myoblasts and cardiac myocytes of rats in vitro, and to study the formation of its syncytia. METHODS: Newborn rats, aged 1 - 3 days, were selected in the study. Myoblasts and cardiac myocytes were identified and purified in vitro, and were inculated to the culture medium together with the same cell number (with the concentration of 1 × 108 L-1 ) . At the 4th day of co-culture, the cells were collected and observed under electron microscopy, they were studied by microinjection of activating biocytin, Texas red labeled avidin was taken as the second antibody, and was observed under fluorescence microscope.
RESULTS: There were 95% anti α-actin cells in myoblasts, desmin im-munochemistry staining was positive. Desmosomes connection but no gap junctions were found between the two kinds of cells by electron microscopy, while gap junction was found from cadiac myocytes aside when biocytin was injected to myotube.
CONCLUSION: In vitro, we find electromechanical coupling can be formed by the connection of myoblasts and cadiac myocytes, which can provide a study method and strong evidence for the syncytia formation of the transplantation of myoblasts into heart.
出处
《中国临床康复》
CSCD
2004年第9期1658-1659,共2页
Chinese Journal of Clinical Rehabilitation
