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

Active Publication Date: 2019-07-02
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

[0010] However, in the prior art, the methods for isolating, purifying and culturing hippocampal neurons of mice and rats are not suitable for naked mole rats. At present, there are no relevant reports on the methods of isolating, purifying and culturing hippocampal neurons of naked mole rats in domestic and foreign literature.

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  • A method for culturing hippocampal neurons of naked mole rats
  • A method for culturing hippocampal neurons of naked mole rats
  • A method for culturing hippocampal neurons of naked mole rats

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention relates to the technical field of cell biology, in particular to a separation, purification and culture method of naked mole rat hippocampal neurons. The hippocampal neurons are separated from newly-born naked mole rat cerebral cortices and purified and cultured by the aid of multiple culture media comprehensively, and a reasonable culture method suitable for poikilothermal rodent mammal naked mole rat hippocampal neurons is found out. With the adoption of the method, a large number of naked mole rat hippocampal neurons with normal functional activity can be acquired conveniently, efficiently and economically, the cells can still keep biological characteristic in the in-vitro state in the in-vitro environment through culture under the low-oxygen condition, so that special physiological functions of the naked mole rat hippocampal neurons are further researched in pure in-vitro cell culture models directly and conveniently, and the important theoretical basis is provided for exploration of the biological mechanism and application to related clinical fields.

Description

[0001] Technical field: [0002] 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 hippocampal neurons of naked mole rats. [0003] Background technique: [0004] Spatial cognition, learning and memory ability is one of the important abilities necessary for the survival of human beings and other animals. The hippocampus is an important part of the limbic system, which plays a pivotal role in various functions of the body, especially in the process of memory acquisition and integration, learning ability acquisition and spatial navigation (Memory systems of the brain: a brief history and current perspective. Squire LR. Neurobiol LearnMem. 2004Nov; 82(3): 171-7.; Place cells, grid cells, and the brain's spatial representation system. Moser EI, Kropff E, Moser MB Annu Rev Neurosci. 2008; 31( ):69-89.). Studies have found that a var...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0793
CPCC12N5/0619C12N2500/34C12N2500/40C12N2500/84C12N2501/13
Inventor 崔淑芳杨文静孙伟汤球丛薇余琛琳林丽芳徐晨蔡丽萍李周桐
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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