Cultivation Of Sheep Skeletal Muscle Satellite Cells And Analysis Of MRFs Gene Expression

Skeletal muscle satellite cells are a kind of myogenic stem cells with proliferative capacity and differentiation potential.It exists between the muscle cell membrane and the basal membrane.These cells play an important role in the growth and regeneration of skeletal muscle.(3 months old)and newly born(0 month old)and middle age sheep(6 months old)were collected from Xilinguole League of Inner Mongolia.The two-step digestion method and tissue mass method The cells were cultured in vitro with 1%gelatin and 4%paraformaldehyde.The adherence time of the cells was measured by means of differential adherence method,percoll density gradient centrifugation and modification The expression of MRFs in skeletal muscle cells was detected by real-time fluorescence quantitative technique.The expression of MRFs in skeletal muscle cells was detected by immunohistochemistry.Draw the following conclusions:The number of cells obtained by the separation and modification of the tissue method was better and the morphology was better and the cells were more pure.A set of highly stable and stable skeletal muscle satellite cells were isolated and purified,system.The cells were identified with desmin and myosine antibodies,and the cytoplasm in the cytoplasm was stained brown or brown,confirming that the cells were skeletal muscle satellite cells.MyoD and Myf5 in the MRFs family are mainly expressed in the proliferative phase of skeletal muscle satellite cells,and MyoG and Myf6 are mainly expressed in the cell differentiation stage.The expression of MyoD and Myf5 increased at the time of cell proliferation,while the expression level of MyoD and Myf6 decreased at the time of differentiation.MyoG and Myf6 were the opposite,and the expression level of MyoD and Myf5 decreased at the time of differentiation.