A method for culturing hippocampal neurons of naked mole rats
A hippocampal neuron and culture method technology, applied in the field of separation, purification and culture of naked mole rat hippocampal neurons, can solve problems such as inapplicability to naked mole rats, achieve good functional state and vitality, simple operation method and high repeatability sexual effect
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Embodiment 1
[0042] Example 1: Isolation, purification and culture of naked mole rat hippocampal neurons
[0043] 1. Experimental materials
[0044] Clean-grade naked mole rats at 60-65 days of pregnancy were provided by the Experimental Animal Center of the Second Military Medical University of the Chinese People's Liberation Army.
[0045] DNase I 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 medium, N2 serum-free supplement factor, etc. were purchased from Thermo Fisher Scientific Company, and 24-well culture plates were purchased from Corning Company.
[0046] The components of the mixed nerve cell culture medium are low-sugar DMEM medium containing 10% fetal bovine serum by volume fraction, and the purification medium for hippocampal neurons is Neurobasal + 2% N2 by volume fraction + 10 μM 5-fluor...
Embodiment 2
[0051] Example 2: Identification of naked mole rat hippocampal neurons
[0052] Identification of the hippocampal neurons of naked mole rats obtained in Example 1: the hippocampal neurons in the serum-free medium were fixed with 4% paraformaldehyde, and identified in combination with morphological identification and immunorefinement chemistry.
[0053] 1. Cell morphology identification:
[0054] Identification methods described in references (Lei Xianga, Yanping Ren, Xun Li, Wen Zhao, YijunSong. MicroRNA-204 suppresses epileptiform discharges through regulating TrkB-ERK1 / 2-CREB signaling in cultured hippocampal neurons. Brain Res. 2016, pii:S0006 -8993(16)30104-4.).
[0055] The result is as figure 1 As shown, under the light microscope, the naked mole-rat hippocampal neurons have larger cell bodies, one or several shorter dendrites, and one elongated axon at the other end.
[0056] 2. Immunocytochemical identification:
[0057] Identification methods described in referenc...
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