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Naked mole rat cortical neuron culture method
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A naked mole rat cortex and culture method technology, applied in the field of cell biology, can solve the problem of inappropriate naked mole rat, etc., and achieve the effects of good functional state and vitality, high repeatability, and simple operation method
Active Publication Date: 2016-06-22
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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[0007] However, in the prior art, the methods for isolating, purifying and culturing cortical neurons of mice and rats are not suitable for naked mole rats. At present, there is no relevant report on the methods of isolating, purifying and culturing cortical neurons of naked mole rats in domestic and foreign literature.
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Embodiment 1
[0036] Example 1: Isolation, purification and culture of naked mole rat cortical neurons
[0037] 1. Experimental materials
[0038] Clean-grade naked mole rats at 65-70 days of pregnancy were provided by the Experimental Animal Center of the Second Military Medical University of the Chinese People's Liberation Army.
[0039] DNase was purchased from Solarbio Biotechnology Co., Ltd., NGF, 5-fluorouracil, trypsin, L-polylysine, penicillin and streptomycin mixture were purchased from Sigma Company, DMEM, low-sugar DMEM, fetal bovine serum from Australia, Neurobasal Culture medium, B27 serum-free additive factors, etc. were purchased from Thermo Fisher Scientific Company, and culture dishes, T25 and T75 culture flasks were purchased from Corning Company.
[0051] Identification of naked mole-rat cortical neurons obtained in Example 1: 4% paraformaldehyde was used to fix cortical neurons in serum-free medium, and combined with morphological identification, immunorefinement chemistry and other methods for identification.
[0053] 鉴定方法参考文献中所述(XuejunChai,ShantingZhao,LiFan,WeiZhang,XiLu,HongShao,ShaoboWang,LingzhenSong,AntonioVirgilioFailla,BerndZobiak,HansG.Mannherz,MichaelFrotscher,Reelinandcofilincooperateduringthemigrationofcorticalneurons:aquantitativemorphologicalanalysis,Development,2016,143:1029-1040)结果如 figure 1 As shown, the naked mole-rat cortex neuron has a large cell body with one or several short dendrites and a long and thin axon at the other end.
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Abstract
The invention relates to the technical field of cellbiology and in particular relates to a separation, purification and culture method of naked mole ratcortical neurons. The cortical neurons 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 rodentmammalnaked mole ratcortical neurons is explored. The method provided by the invention can be used for simply, conveniently, efficiently and economically obtaining a lot of naked mole rat cortical neurons 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 cortical neurons can be conveniently and directly researched in a pure in-vitro cellculture 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 cellbiology, in particular to a method for separating, purifying and culturing cells, in particular to a method for separating, purifying and culturing naked mole rat cortical neurons. Background technique: [0002] Although neurons do not have the highest proportion in the mammalian central nervous system, they are the most functionally important type of cells. Neurons play a vital role in synapse formation, synaptic signal transmission, and synaptic plasticity . The neurons in the cerebral cortex play the role of the control center in many functions of the body, including vision, hearing, smell, language, movement, and body surfacesensation. [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%. Naked mole rats live in dark and na...
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IPC IPC(8): C12N5/0793
CPCC12N5/0619C12N2500/40C12N2501/13
Inventor 崔淑芳杨文静孙伟汤球肖邦赵善民丛薇程继帅
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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