Separation and culture method of rat Schwann cells

Abstract

The invention provides a method for isolating and culturing rat Schwann cells, comprising the following steps: (1) disinfecting tissue processing instruments; (2) selecting 2-3 week old male SD rats, killing them by neck dislocation, and using ophthalmic scissors Cut off the hair on the legs of rats, cut off the sciatic nerve, put the tissue into pre-cooled sterile PBS solution containing double antibodies and wash several times to remove blood stains and adventitia; (3) Mince the tissue, and use preheated type II collagen (4) Terminate the digestion, centrifuge, discard the supernatant, add culture medium and resuspend; (5) Inoculate the tissue block into a 6-well culture plate for culture by adherent method; (6) Incubate for 1-2 hours, Placed in a 37°C, 5% CO2 environment for culture; (7) Cell subculture and purification; (8) Cell morphology observation and identification. The method for separating and culturing rat Schwann cells provided by the present invention is simple and easy to operate, stable and efficient, and the obtained Schwann cells have high vigor, which can be used to establish model cells for in vitro experiments and meet the requirements of rat Schwann cell experiments .

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 rat Schwann cells. Background technique [0002] Schwann cells (Schwann cells SCs) are the unique and most important glial cells in the peripheral nervous system, which play an important role in the occurrence and regeneration of peripheral nerves after injury. It is the sheath cells of peripheral nerve fibers, arranged in strings, wrapping the axons of peripheral nerve fibers, and wrapping repeatedly to form concentric circular lamellar layers, forming myelin sheath. Schwann cells can secrete cell adhesion molecules, a variety of neurotrophic factors (NTFs) and extracellular matrix to promote and guide the regeneration of peripheral nerve axons. [0003] Schwann cells can continuously secrete a large number of growth factors VEGF, TGF, PDGF and other biologically active substances as well as secretio...

AI Technical Summary

Problems solved by technology

At present, there are few studies on the isolation and culture of rat Schwann cells at home and abroad, the isolation method is immature, and the obtained cells are of low purity and small number.

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