原代咬肌肌卫星细胞的提取及鉴定

Isolation and Differentiation of Primary Muscle Satellite Cell from Rat Masseter.

目的:探讨原代咬肌肌卫星细胞的提取、培养、鉴定的方法及其生物学特性。方法:采用链霉蛋白酶消化法和Percoll非连续密度离心法获取大鼠咬肌肌卫星细胞,进行体外原代培养。通过免疫荧光染色鉴定原代咬肌肌卫星细胞激活的状况和肌管的融合情况。结果:采用链霉蛋白酶消化法和Percoll非连续密度离心法可获得纯度较高的肌卫星细胞。体外培养初期,运用免疫荧光染色鉴定Pax7和MyoD,大部分的咬肌肌卫星细胞处于激活状态。在分化培养基条件下,咬肌肌卫星细胞可分化融合形成肌管,MyHC染色呈阳性。结论:原代咬肌肌卫星细胞可被分离并体外培养,具有良好的分化能力。

Objective.. To establish the methods of isolation, culture, identification, and biological characteristics of primary muscle satellite cell from rat masseter （MAS--derived satellite Cells）. Methods.. We used Pronase and Percoll gradient centrifugation to isolate and pu.rify satellite cell. Using Immunostaining,we can identify the activa tion of MAS--derived satellite cells and myotube formation. Results： Through Pronase and Percoll gradient centrif ugation, we can get highly purified primary MAS--derived satellite ceils. Using Immunostaining for Pax7 and MyoD, we found most primary satellite cell were activated and did not make a decision to self--renew or differenti- ate at the early stage（24h）. Immunostaining for MyHC was to observe the myotube formation. Conclusion： Through Pronase and Percoll gradient centrifugation, we can get highly purified primary MAS-- derived satellite cells and these primary satellite cell can differentiate and form myotube well.