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Naked mole rat oligodendroglia precursor cell culture method

A technology of precursor cells and culturing methods, which is applied in the field of separation, purification and cultivation of oligodendrocyte precursor cells of naked mole rats, can solve the problem of inapplicability to naked mole rats, and achieves the maintenance of cell viability and function and high reproducibility. The effect of sex, simple operation method

Active Publication Date: 2016-06-22
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the prior art, the isolation, purification and culture methods of oligodendrocyte precursor cells in mice and rats are not suitable for naked mole rats. Related reports on purification and culture methods

Method used

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  • Naked mole rat oligodendroglia precursor cell culture method
  • Naked mole rat oligodendroglia precursor cell culture method
  • Naked mole rat oligodendroglia precursor cell culture method

Examples

Experimental program
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Effect test

Embodiment 1

[0041] Example 1: Isolation, purification and culture of naked mole rat oligodendrocyte precursor cells

[0042] 1. Experimental materials

[0043] Clean-grade naked mole rats born 1-7 years old were provided by the Experimental Animal Center of the Second Military Medical University of the Chinese People's Liberation Army. Place the newborn naked mole rats on an ice plate at -20°C for anesthesia, soak them in 75% ethanol for 10 minutes, then decapitate them, and wipe the surface of the naked mole rats' heads with a mixture of penicillin 100 U / ml and 0.1 mg / ml streptomycin , then cut the head skin and skull, peel off the brain and place it in pre-cooled dissecting fluid.

[0044] DNase I was purchased from Solarbio Biotechnology Co., Ltd., trypsin, L-polylysine, penicillin-streptomycin mixture, etc. were purchased from Sigma Company, DMEM, low-sugar DMEM, fetal bovine serum from Australia, Neurobasal medium, B27-free Serum added factors, etc. were purchased from Thermo Fishe...

Embodiment 2

[0051] Example 2: Identification of naked mole rat oligodendrocyte precursor cells

[0052] The identification of the naked mole rat oligodendrocyte precursor cells obtained in Example 1: the oligodendrocyte precursor cells in the proliferation medium were fixed with 4% paraformaldehyde, and combined with morphological identification, immune cell identified by chemical methods.

[0053] 1. Cell morphology identification:

[0054] The identification method is described in references (WenjingYang, LinXiao, CuiLi, XiuyunLiu, MingdongLiu, QiShao, DanWang, AijunHuang and ChengHe. TIP30inhibitsoligodendrocyteprecursorcelldifferentiationviacytoplasmicssequestrationofOlig1.Glia.2015;63(4):684-98.).

[0055] The result is as figure 2 As shown, under the light microscope, the cell body of the oligodendrocyte precursor cell is round or oval, the cell body has a strong refractive index, and there is an obvious halo around the cell body, and dipolar or tripolar protrusions protrude from...

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Abstract

The invention relates to the technical field of cell biology and in particular relates to a separation, purification and culture method of naked mole rat oligodendroglia precursor cells. The oligodendroglia precursor cells are separated from cerebral cortices of naked mole rat fetal rats and are purified and cultured by comprehensively utilizing a plurality of types of culture mediums, and a reasonable culture method applicable to poikilothermal rodent mammal naked mole rat oligodendroglia precursor cells is explored. The method provided by the invention can be used for simply, conveniently, efficiently and economically obtaining a lot of naked mole rat oligodendroglia precursor cells with normal functional activity, and cells can still keep biological characteristics under an in-vivo state in an in-vitro environment through culture under a low-oxygen condition, so that special physiological functions of the naked mole rat oligodendroglia precursor cells can be conveniently and directly researched in a pure in-vitro cell culture model, and furthermore, an important theoretical basis is provided for exploring a biological mechanism and applying the biological mechanism to clinically relative fields.

Description

Technical field: [0001] The invention relates to the technical field of cell biology, in particular to a cell separation, purification and culture method, in particular to a naked mole rat oligodendrocyte precursor cell separation, purification and culture method. Background technique: [0002] Oligodendrocytes are myelinating cells of the central nervous system, and the myelin sheath structure formed around the axons of myelinated neurons has the functions of nourishing, supporting and protecting neurons, and the existence of myelin sheath is the key to neurons. The structural basis for the efficient conduction of action potentials in axons can ensure the speed of information transmission between neurons. Oligodendrocytes are differentiated from oligodendrocyte precursor cells. Under pathological conditions, oligodendrocyte precursor cells can be stimulated by the microenvironment to initiate differentiation again. In many diseases related to cerebral ischemia and hypoxia,...

Claims

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Application Information

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IPC IPC(8): C12N5/079
Inventor 崔淑芳杨文静孙伟汤球
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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