Establishment of the methods for isolation,culture and differentiation induction of satellite cells from bovine skeletal muscle

To further reveal the mechanisms of muscle differentiation and provide important help for transgenic cattle research.The skeletal muscles obtained from new-born fetal bovine were used as materials.Collagenase Ⅰ and Ⅺ were combined with trypsin for the digestion,respectively.The skeletal muscle satellite cells were isolated and purified by the differential adhesion method,and then they were also identified using immunofluorescence staining,RT-PCR and Western-blot.The results showed that a large number of bovine skeletal muscle satellite cells were obtained successfully.The purity for mRNA and protein expression of the cell markers was more than 98%.The cells grew in good condition and could passage 90 generations to maintain strong proliferative activity.The cells kept high differentiation efficiency,the cells were fused to form polynuclear myotubes using 2% horse serum to induce the cellular differentiation.Many myotubes could fuse to thicker myotubes spontaneously,and the shrinkage could be observed.