Mouse skeletal muscle satellite cell separation and culture method

Abstract

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.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and in particular relates to a method for separating and culturing mouse skeletal muscle satellite cells. Background technique [0002] Skeletal muscle satellite cells (skeletal muscle satellite cells) are myogenic stem cells with differentiation and proliferation potential in skeletal muscle. They originate from the embryonic mesoderm and distribute in the shape of a small mononuclear spindle between the muscle fiber serosa and the basement membrane. Under normal conditions In the resting state, when the muscle is stimulated by injury, the cells undergo mitosis. The new cells can form muscle bundles and repair muscle tissue damage. Muscle satellite cells are a new hotspot in the research of genetic engineering and embryonic development. To establish a three-dimensional injury model for further research on the mechanisms of muscle injury and repair and regeneration....

AI Technical Summary

Problems solved by technology

[0004] The cells isolated by the tissue block adherence method are not high in purity, the number is small, and the culture time is long

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