Primary Culture And Identification Of Mice Muscle Satellite Cell

Objective: To establish the reliable method of primary culture and identification of mice muscle satellite cell in vitro.Methods: Mice muscle satellite cell were obtained from posterior limb skeletal muscle. Then made cell suspension. The cells were separated by Percoll method and purified with different speed adherence method. Cells were observed under inverted microscope and then drawed cell growthing curve. Satellite cells were identified with monoclonal antibody against desmin and α-actin. Results: Primary muscle satellite cells were round before differentiation. The purity of satellite cells was more than 80%. Satellite cells grew well in F10 cultural medium. Cells began to adherence the wall gradually after 1 day. The cells were small and round. Cells grew fastly in 3 to 6 day. 50%～70% dish bottom was covered with single layer cells after 7-10 days. Satellite cell proliferation was slow after 12-14 days. Desmin and actin immunocytochemical stain was clear between 10-15 days.Conclusion: High purity muscle satellite cells could be obtained by Percoll separating and different speed adherencing method. Immunocytochemical detection of desmin and actin is the effective method to determine muscle satellite cell.