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Method for separating and purifying oligodendrocyte precursor cells

A precursor cell, separation and purification technology, applied in the field of separation and purification of oligodendrocyte precursor cells, can solve the problems of low acquisition rate of OPCs, cell death, long time consumption, etc., and achieve good proliferation growth and directional differentiation ability, High-efficiency, high-purity, actionable effects

Inactive Publication Date: 2010-06-16
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have the following problems: ①The operation steps are not easy to master; ②It takes a long time, generally 18-20 days; ③The acquisition rate of OPCs is low, and each newborn rat can obtain about 10 4 OPCs; ④The purity of OPCs is low, usually 90% to 95%; ⑤Mechanical separation causes great damage to cells and easily causes a large number of cell death; ⑥Stimulation of multiple cell growth factors is required, and the cost is high

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  • Method for separating and purifying oligodendrocyte precursor cells
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  • Method for separating and purifying oligodendrocyte precursor cells

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Embodiment Construction

[0026] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

[0027] 1. Experimental materials

[0028] Clean-grade SD rats born 0 to 3 days old were purchased from the Animal Center of the Third Military Medical University of the Chinese People’s Liberation Army. The rats were anesthetized by burying them in ice for 5 minutes, their heads were wiped with alcohol, and their heads were decapitated. Bacteria were washed twice in PBS to remove blood, and then placed in sterile D-Hank's solution containing penicillin at a concentration of 160 U / ml and streptomycin at a concentration of 200 U / ml at a temperature of 4°C, and stripped under a dissecting microscope. Brain, spare.

[0029] Trypsin was purchased from Boster Company, DNase was purchased from Shanghai Sangon Bioengineering Technology Service Co., Ltd., Poly-D-Lysine (Poly-D-Lysine), insulin and triiodothyronine (Triiodothyronine) were purchas...

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Abstract

The invention discloses a method for separating and purifying oligodendrocyte precursor cells. The method comprises: digesting cerebral cortex of neonatal rats and part of white matter with trypsin and DNA enzyme; blowing and beating the obtained product to be a single-cell suspension; inoculating the single-cell suspension into a culture flask coated with polylysine in advance by use of a mixed-cell culture medium; performing culture for 3 to 5 days; replacing the culture flask with an oligodendrocyte precursor cell proliferation culture medium when the fusion of bottom-layer cells reaches 65 to 75 percent; performing culture for 3 to 5 days; replacing the oligodendrocyte precursor cell proliferation culture medium with an oligodendrocyte precursor cell separation culture medium; digesting the obtained product at 37 DEG C; isolating oligodendrocyte precursor cells from mixed cells; blowing and beating the obtained product to be the single-cell suspension; inoculating the single-cell suspension into the culture flask coated with polylysine in advance by use of an oligodendrocyte precursor cell inoculation culture medium; replacing the culture flask with an oligodendrocyte precursor cell purification culture medium after the cells adhere to a wall; performing culture for 2 to 4 days; and obtaining adherence cells, namely the oligodendrocyte precursor cells. The method can obtain a large number of high-purity good-activity oligodendrocyte precursor cells conveniently, rapidly, economically and efficiently.

Description

technical field [0001] The invention relates to a method for separating and purifying cells, in particular to a method for separating and purifying oligodendrocyte precursor cells. Background technique [0002] Oligodendrocytes are the myelin-forming cells of the central nervous system, which play a crucial role in facilitating the transmission of nerve impulses and maintaining the survival of nerve axons. More and more studies have shown that many mental and neurological diseases such as schizophrenia, depression, multiple sclerosis, metachromatic leukoencephalopathy, traumatic spinal cord injury recovery disorders, etc. are related to the development of oligodendrocytes. Abnormalities, demyelination or dysmyelination. Therefore, understanding the biological characteristics of oligodendrocytes has guiding significance for the treatment of oligodendrocyte-related diseases. [0003] At present, the research on oligodendrocyte-related diseases mostly adopts the in vivo demye...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
Inventor 牛建钦肖岚
Owner ARMY MEDICAL UNIV
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