a simplified but robust method for the isolation of avian and mammalian muscle satellite cells一个简单但强大的方法隔离的鸟类和哺乳动物的肌肉卫星细胞.pdfVIP

A simplified but robust method for the isolation of avian and mammalian muscle satellite cells Baquero-Perez et al. Baquero-Perez et al. BMC Cell Biology 2012, 13:16 /1471-2121/13/16 21 June 2012 Baquero-Perez et al. BMC Cell Biology 2012, 13:16 /1471-2121/13/16 METHODOLOGY ARTICLE Open Access A simplified but robust method for the isolation of avian and mammalian muscle satellite cells * † † Belinda Baquero-Perez , Suresh V Kuchipudi , Rahul K Nelli and Kin-Chow Chang Abstract Background: Current methods of isolation of muscle satellite cells from different animal species are highly variable making inter-species comparisons problematic. This variation mainly stems from the use of different proteolytic enzymes to release the satellite cells from the muscle tissue (sometimes a single enzyme is used but often a combination of enzymes is preferred) and the different extracellular matrix proteins used to coat culture ware. In addition, isolation of satellite cells is frequently laborious and sometimes may require pre-plating of the cell preparation on uncoated flasks or Percoll centrifugation to remove contaminating fibroblasts. The methodology employed to isolate and culture satellite cells in vitro can critically determine the fusion of myoblasts into multi- nucleated myotubes. These terminally differentiated myotubes resemble mature myofibres in the muscle tissue in vivo, therefore optimal fusion is a keystone of in vitro muscle culture. Hence, a simple method of muscle satellite cell isolation and culture of different vertebrate species that can result in a high fusion rate is highly d