Method for separating and purifying oligodendrocyte precursor cells

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...

AI Technical Summary

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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