大鼠成肌细胞体外培养及其自发收缩特性
免疫组织化学,,成肌细胞;细胞培养;收缩;免疫组织化学,0引言,1材料和方法,2结果,3讨论,【参考文献】
In vitro culture and autocontraction of SpragueDawley rats’ skeletal myoblastsZHAO KeYan, YU ShiQiang, CAI ZhenJie, WU TieJun, HU Jun, ZHOU HePing
Department of Cardiovascular Surgery, Xijing Hospital, Fourth Military Medical University, Xi’an 710033, China
【Abstract】 AIM: To establish a practical procedure for culturing SpragueDawley rats’ myoblasts in vitro and to describe the characteristics of these myoblasts. METHODS: Muscle samples were minced and digested by collagenaseⅠand purified with Percoll gradient liquid. Myoblasts were tested by αactin and desmin immunochemistry stains. Many of the myoblasts could autocontract, so isoproterenol (IP) was added into the culture bottles with the final density as 0.1μmol/L so as to compare the contracting rates. RESULTS: The myoblast viability was measured as up to 90 % by trypan blue. Cell proliferation reached the peak 3-5 d after the cells were planted, and cell purity could be as high as 95 % by αactin and desmin immunochemistry stains tests. Myoblastes could contract on the 2nd or 3rd d after they were fused and the contracting rates ranged from 50 to 160 beats per minute for the most part, while some contracted with 200 beats per minute. When isoproterenol was added, the mean contracting rate increased about 18.4 beats per minute compared with that before the adding isoproterenol. CONCLUSION: Myoblasts can autocontract and IP can increase their contracting rates. The transplant of myoblasts into the ischemia heart has been proved to be effective in the treatment of heart diseases. ......
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