A method for culturing naked mole rat astrocytes

An astrocyte and culture method technology, which is applied in the field of separation, purification and culture of naked mole rat astrocytes, can solve the problem of inapplicability to naked mole rat, etc., and achieves high repeatability and simple operation method. Effect

Active Publication Date: 2019-09-27
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, in the prior art, neither mouse nor rat astrocyte isolation, purification and culture methods are suitable for naked mole rats. At present, there is no method for the separation, purification and culture of naked mole rat astrocytes in the literature at home and abroad. related reports

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  • A method for culturing naked mole rat astrocytes
  • A method for culturing naked mole rat astrocytes
  • A method for culturing naked mole rat astrocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Isolation, purification and culture of naked mole rat astrocytes

[0038] 1. Experimental materials

[0039]Clean-grade naked mole rats born 1-9 days 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.

[0040] DNase I was purchased from Solarbio Biotechnology Co., Ltd., trypsin, penicillin-streptomycin mixture, etc. were purchased from Sigma Company, DMEM, low-sugar DMEM, fetal bovine serum from Australia, Neurobasal medium, N2 serum-free additive factors, etc. were purchased from Thermo Fisher Scientific company, petri dishes,...

Embodiment 2

[0046] Example 2: Identification of naked mole rat astrocytes

[0047] Identification of naked mole-rat astrocytes obtained in Example 1: 4% paraformaldehyde was used to fix astrocytes in serum-free medium, combined with morphological identification, immunorefinement chemistry, etc. method for identification.

[0048] 1. Cell morphology identification:

[0049] The identification method was described in the literature (Chung WS, Clarke LE, Wang GX, Stafford BK, Sher A, Chakraborty C, Joung J, Foo LC, Thompson A, Chen C, Smith SJ, Barres BA.Astrocytesmediate synapse elimination through MEGF10 and MERTK pathways. Nature, 2013, 504(7480): 394-400.).

[0050] The result is as figure 2 As shown, under the light microscope, the astrocyte cell body and nucleus are larger, the cytoplasm volume is also larger, the refractive index of the cell body is poor, and irregular pseudopodia-like protrusions protrude from the cell body to the surrounding.

[0051] 2. Immunocytochemical iden...

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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 astroglia cells. The astroglia cells are separated from cerebral cortices of newly-born naked mole 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 astroglia 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 astroglia 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 astroglia 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 method for separating, purifying and culturing cells, in particular to a method for separating, purifying and culturing naked mole-rat astrocytes. Background technique: [0002] Astrocytes are the most abundant type of cells in the mammalian central nervous system. As more and more studies have found that astrocytes play a role in synapse formation, synaptic signal transmission efficiency, and synaptic plasticity, astrocytes have gradually attracted more attention from researchers. [0003] The naked mole rat is a temperature-changing mammal. It can maintain its brain structure and function intact and carry out normal life activities in an unventilated cave with an oxygen concentration of only about 6%. Studies have found that astrocytes often extend pseudopods to attach around the capillaries in the brain, so as to closely sense changes in oxygen concentration and ion e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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