A kind of naked mole rat oligodendrocyte precursor cell culture method
A technology of precursor cells and culture methods, which is applied in the field of separation, purification and culture of naked mole rat oligodendrocyte precursor cells, can solve the problems of being unsuitable for naked mole rats, maintain cell viability and function, and have simple operation methods , Highly reproducible results
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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 was described in references (Wenjing Yang, Lin Xiao, Cui Li, Xiuyun Liu, Mingdong Liu, Qi Shao, Dan Wang, Aijun Huang and Cheng He. TIP30 inhibitsoligodendrocyte precursor cell differentiation via cytoplasmic sequestration of Olig1.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 protr...
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