Study on culture, identification and differentiation of primary rat skeletal muscle satellite cells

Abstract

To establish the isolation, culture and identification methods of primary rat skeletal muscle satellite cells (SMSC) and observe its characterization of differentiation in vitro. Skeletal muscle satellite cells were obtained by tissue block culture method in combination with pre-plating techniques, and the purity of these cells was detected by both immunocytochemistry and fluorescence activated cell sorter (FACS) with Pax7 as marker of SMSC. Myogenesis of these cells was induced in differentiation medium and the mRNA expressions of myogenic differentiation gene (MyoD) and Myogenin were determined by Real-time polymerase chain reaction (PCR). Cells crawled out from the edge of tissue blocks after 1 week of culture. After purification by pre-plating techniques, more than 97.6% of the cells expressed Pax7, a unique marker of satellite cells. The mRNA of MyoD and Myogenin showed time-specific expression in the myogenesis induction process in vitro. Skeletal muscle satellite cells with high purity and strong differentiation ability can be obtained by means of tissue block culture method.