The invention provides a mouse
skeletal muscle satellite cell separation and culture method. The mouse
skeletal muscle satellite cell separation and culture method comprises following steps: 1, mice are killed through
intraperitoneal injection of pentobarbital
sodium, and are immersed in 75%
alcohol for disinfection; 2, the mice are fixed on plates with the venter posterior surfaces upward, hind limb
skin is peeled under aseptic conditions, and thighbone
muscle tissues are collected; 3, in an aseptic operation bench, PBS washing is carried out for three times, surface blood vessels, tendons, and connective tissues are removed; 4, the processed tissues are
cut into pieces, and are subjected to
digestion with preheated
trypsin and IV type
collagenase respectively; 5,
digestion is stopped,
centrifugation is carried out, and an obtained supernatant is removed; 6, a
cell filtrate is collected, and trypan-blue
drying detection is adopted to detect
cell activity; 7, after
cell counting, a complete medium is adopted for resuspension
inoculation into a coated culture flask for culture at 37 DEG C at 5% CO2 environment; 8, differential attachment method is adopted to remove fibrocytes; 9,
cell morphology is carried out; 10,
ELISA method detection is carried out; and 11, RT-PCR detection is carried out. The mouse
skeletal muscle satellite cell separation and culture method is simple in operation, and is high in
survival rate; the number of obtained cells is large; the mouse skeletal
muscle satellite
cell separation and culture method is an ideal method, and is capable of providing experiments with reliable cell resources.