Method for separating and culturing mouse synovial cells

A technology of synovial cells and a culture method, applied in the field of cell culture of modern biotechnology, can solve the problems of long culture time and low cell purity, and achieve the effects of easy preservation, reliable cell resources and good activity

Inactive Publication Date: 2017-12-15
JIANGYIN CHI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Most of the established mouse synoviocyte isolation and culture methods are based on enzymatic digestion, and the purity of the isolated cells is not high, and the culture time is long

Method used

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  • Method for separating and culturing mouse synovial cells
  • Method for separating and culturing mouse synovial cells
  • Method for separating and culturing mouse synovial cells

Examples

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Embodiment Construction

[0037] The terms used in the present invention, unless otherwise specified, generally have the meanings commonly understood by those skilled in the art.

[0038] The present invention will be described in detail below in conjunction with the accompanying drawings and examples, and the protection content of the present invention is not limited to the following examples.

[0039] In the following examples, various procedures and methods not described in detail are conventional methods well known in the art.

[0040] The experimental instruments and reagents used in this experiment are as follows:

[0041] A set of surgical instruments, inverted microscope (XDS-1A, Shanghai), fluorescence microscope (Leica, USA), cryogenic centrifuge (TD24B-WS, Shanghai), ultra-low temperature refrigerator (Zhongke Meiling), pipette gun (Eppendorf, USA) ), electronic analytical balance (Sartorius, USA), ultra-clean bench (HJ-CJ-1D, Shanghai), CO 2 Cell culture incubator (SANYO MCO-17AI, Japan)....

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Abstract

The invention provides a method for separating and culturing mouse cells, comprising the following steps: (1) disinfecting tissue processing instruments; (2) selecting male BALB/c mice aged 1 to 2 months, killing them by dislocation of the neck, and using surgical scissors Separate the synovial layer and fibrous layer of the joint capsule, then take out the synovial layer tissue, put the tissue into pre-cooled sterile double-antibody-containing PBS solution and wash several times to remove blood stains and fat tissue; (3) Cut the tissue into pieces and use Digest with preheated type II collagenase and trypsin respectively; (4) terminate the digestion, centrifuge, and discard the supernatant; (5) inoculate into the coated culture flask by artificial adherence method; (6) after incubation for 1-2 hours, Cultured in an environment of 37°C and 5% CO2; (7) observation of cell morphology; (8) subculture of cells; (9) identification of cells. The method for isolating and culturing mouse synoviocytes provided by the invention is simple to operate, has a large number of cells and a high survival rate, and is an ideal primary method for isolating and culturing mouse synoviocytes, providing a reliable basis for experiments. cellular resources.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, in particular to a method for separating and culturing mouse synoviocytes. Background technique [0002] The synovium (synovium) is the inner layer of the joint capsule, which is light red, thin and soft, and is composed of loose connective tissue. The synovial membrane secretes synovial fluid, which plays an important role in joint movement. The normal synovium is divided into two layers, a thin cellular layer (luminal layer) and a vascular layer (subintimal layer). [0003] Synoviocytes originate from the mesenchyme in the skeletal germ base. Synovial tissue contains a variety of cells, such as synoviocytes, leukocytes, fat cells, etc. There are two types of synoviocytes, type A and type B. Macrophage-like type A cells (MLC) have filopodia, have phagocytic function, and do not have the ability to proliferate. B-type synoviocytes (FLC), also known as fibroblast ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0652C12N2501/33C12N2509/00C12N2533/32
Inventor 何刚张亚洲蒋敏齐来俊
Owner JIANGYIN CHI SCI
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