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Method for separating and culturing olfactory ensheathing cells of mice

A technology for the separation and cultivation of olfactory ensheathing cells, which is applied in the field of medicine and biology, can solve the problems of weak biological activity and low density per unit area of ​​olfactory ensheathing cells, and achieve axon extension promotion effect, fast cell proliferation, and large number Effect

Pending Publication Date: 2019-04-26
李海明
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The prior art uses human embryos, male rats or newborn rats to isolate and purify olfactory ensheathing cells, but the density per unit area of ​​isolated and cultured olfactory ensheathing cells is small and the biological activity is not strong

Method used

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  • Method for separating and culturing olfactory ensheathing cells of mice
  • Method for separating and culturing olfactory ensheathing cells of mice
  • Method for separating and culturing olfactory ensheathing cells of mice

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Embodiment Construction

[0022] In order to make the object, technical solution and advantages of the present invention clearer, the following examples further describe the present invention in detail. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0023] Materials and equipment used in the experiment of the present invention include: P75 mouse monoclonal antibody and GFAP mouse monoclonal antibody were purchased from Abcam Company in the United Kingdom; fetal bovine serum was purchased from CLARK Company in the United States, and DF12 medium was purchased from Hyclone Company in the United States. The system was purchased from Nikon, Japan, and the carbon dioxide incubator was purchased from Thermo, USA. The complete medium used in the test process of the present invention is DF12 medium, and the complete medium containing fetal bovine serum is: DF12 medium containing 15% fetal bovine serum.

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PUM

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Abstract

The invention provides a method for separating and culturing olfactory ensheathing cells of mice. The method comprises the steps of taking a female mouse in the middle and late pregnancy, taking out amouse embryo, taking out an olfactory bulb of the mouse embryo, cutting the olfactory bulb tissue into pieces, adding a complete medium containing serum, filtering with a 200-mesh sieve, sucking thetissue fluid out and adding into a centrifuge tube for centrifuging, discarding the supernatant, resuspending, and repeating the operation for three times; adding into the complete medium to adjust the cell concentration, and culturing in a constant temperature incubator; then, using a differential adhension method to purify the olfactory ensheathing cells. After the method for separating and culturing the olfactory ensheathing cells of the mice is adopted, the separated and cultured olfactory ensheathing cells are large in quality, high in density and strong in biological activity.

Description

technical field [0001] The invention relates to the field of medical biology, in particular to a method for isolating and culturing rat olfactory ensheathing cells. Background technique [0002] The olfactory bulb area is different from other parts of the central nervous system. This area has a strong ability to regenerate nerves. The reason is that there is a special glial cell in this area——olfactory ensheathing cells (OECs), which can repair damaged nerves. Tissue repair creates the right environment. The treatment of various nervous system diseases by OECs transplantation has become a hotspot in neuroprosthetics. Obtaining a large amount of OECs with high purity and good biological activity is the key to the success of experimental research and the prerequisite for clinical application. In the prior art, human embryos, male rats or newborn rats are used to isolate and purify olfactory ensheathing cells, but the isolated and cultured olfactory ensheathing cells have a l...

Claims

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Application Information

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IPC IPC(8): C12N5/079
CPCC12N5/0622C12N2509/00C12N2501/06
Inventor 李海明金保哲
Owner 李海明
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