Schwann cell extraction, purification and culture method

A culture method and cell technology, which is applied in the field of extraction, purification and culture of Schwann cells, can solve the problems of poor activity, low nerve acquisition, unfavorable primary culture and expansion of Schwann cells, etc. low volume effect

Active Publication Date: 2017-12-22
SOUTHERN MEDICAL UNIVERSITY
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Problems solved by technology

However, in the prior art, the isolation, purification and culture methods of Schwann cells from mice and rats all have the problems of low nerve acquisition and large residual fi

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  • Schwann cell extraction, purification and culture method
  • Schwann cell extraction, purification and culture method

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[0070] Example 1 A method for extraction, purification and culture of Schwann cells

[0071] (1) Preparation before experiment:

[0072] 1ml of 0.1mg / ml PLL distilled water solution, soak a 35mm petri dish in a cell incubator at 37°C overnight; blot the PLL distilled water solution, rinse with distilled water twice, and dry for 30 minutes under ultraviolet radiation;

[0073] Prepare 50ml HBSS and pre-cool at 4℃;

[0074] Sterilize scissors and tweezers soaked in 75% ethanol for 30 minutes, and then dry under ultraviolet light for 30 minutes;

[0075] Ultraviolet disinfection operation table.

[0076] (2) Separate the skin and muscle: fix the newborn SD rats with 75% ethanol skin disinfection into a specimen shape with the ventral side down (such as figure 1 -A); dissection scissors to separate the back skin (such as figure 1 -B), separate the muscle layer by layer (such as figure 1 -C);

[0077] (3) Cut the brachial plexus and sciatic nerve: first cut the brachial plexus at the distal ne...

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Abstract

The invention discloses Schwann cell extraction, purification and culture method which comprises the following steps: cutting off brachial plexus and ischiadic nerve, further removing spinal cord, and extracting peripheral nerve from different foramen intervertebrales; digesting the peripheral nerve so as to obtain a single-cell suspension; and performing amplification culture, thereby obtaining Schwann cells. By adopting the method disclosed by the invention, the nerve extracted from the foramen intervertebrales is free of epineuria and is low in fibroblast residue. According to the method, the peripheral nerve (including the brachial plexus, the intercostal nerve and the ischiadic nerve) is extracted from the foramen intervertebrales, a nerve amount which is 6-8 times of that of a conventional method is obtained, the residue of fibroblast is remarkably reduced, and thus culture and amplification of the Schwann cells are facilitated.

Description

technical field [0001] The invention relates to a method for extracting, purifying and culturing Schwann cells. Background technique [0002] Primary cultured Schwann cells are not only the basis for studying Schwann cell function and surrounding myelin biology, but also important seed cells for neural tissue engineering. Glial cells in the peripheral nervous system are called Schwann cells, which are distributed along the processes of neurons. Schwann cells wrap around nerve fibers called myelinated nerve fibers. The morphology and function of Schwann cells of myelinated and unmyelinated nerve fibers are different. The outer surface of Schwann cells has a basement membrane, which can secrete neurotrophic factors and promote the survival of damaged neurons and the growth of axons. Regeneration, involved in the formation of nerve fibers in the peripheral nervous system. Immature Schwann cells can be stimulated by the microenvironment to initiate differentiation again. Spi...

Claims

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Application Information

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IPC IPC(8): C12N5/079
CPCC12N5/0622C12N2501/01C12N2501/11C12N2509/10
Inventor 郭家松文锦坤王祥海谭丹丹李莉霞
Owner SOUTHERN MEDICAL UNIVERSITY
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