摘要
目的:探寻共同培养的骨骼肌成肌细胞与心肌细胞间连接的形态学证据。方法:将鼠心肌细胞与骨骼肌成肌细胞体外共培养,于第2、4、7、14天免疫组化染色钙粘素(Cadherin)、细胞间隙连接蛋白73(Connexin43)、神经细胞粘附分子(NCAM)3种蛋白,并行定量RT-PCR检查分组比较分析;于第4、7、10、14天透射电镜观察。结果:共同培养物中的骨骼肌成肌细胞表达Connexin43,且随时间延长而增加;Cadherin表达增加不明显;NCAM含量维持在较低水平,至2周时才表达较多;透射电镜下观察到两者间存在类似于缝隙连接的连接。结论:与心肌细胞共培养后骨骼肌成肌细胞融合、成熟延迟,Connexin43表达增多;两者间确有形成类似于缝隙连接的物质基础并可形成连接。
Objective：To investigate the morphological evidence for the connection of cocultured myoblasts and myocytes. Methods： Rat myocytes and skeletal myoblasts were cocultured with the ratio of 4：1. Cadherin,Connexin43 and NCAM were stained by ABC-AEC staining on day 2,4,7 and 14 respectively. The staining results were observed under microscope. Cells were compare d by transmission electroscope on day 4,7,10 and 14. Three groups were set as followed： experiment group （myocyte and myoblast were co-cultured with ratio 4：1）, control group 1 （myoblast was cultured alone with same cell account）, and control group 2 （myocyte was cultured alone with same cell account）. Cells were digested and gathered on day 2,4,7,14 after cultured and RT-PCR was applied. Results were compared among the 3 groups. Results： Connexin43 was expressed in mypblasts and increased with cultural duration among the cocultured myoblasts and myocytes. The expression of cadherin also increased with the prolongation of co-culture time but less significant than that of connexin43. The expression of NCAM remained low until two weeks after co-culture. Connection similar to gap junction was observed to exist between myoblasts and myocytes under transmission electroscope. Conclusion： The myoblasts myocytelized in some characters after cocultured with myocytes： the fusion and maturation was delayed, the expression of connexin43 was increased Material bases and possibility for the connection similar to gap junction do exist among the co-cultured skeletal myoblast and cardiac myocytes.
出处
《中国临床医学》
2009年第2期180-183,共4页
Chinese Journal of Clinical Medicine
基金
上海市重点学科建设项目(B116)
