Method for culturing primary hippocampal neurons

Abstract

The invention discloses a method for cultivating primary hippocampal neurons. The method comprises: (1) digesting the hippocampal tissue; (2) blowing and dispersing the digested hippocampal tissue to obtain a cell suspension; (3) The cell suspension is inoculated and first cultivated for 2-4 hours to make the cells adhere to the wall; (4) the first culture solution in the culture dish is replaced with the second culture solution, and the second culture is carried out for 24 hours; (5) to Add 1-2 μg / ml of cytarabine to the culture dish, and carry out the third culture for 24 hours; (6) Use the second culture medium to replace half of the medium of the culture dish, and carry out the fourth culture for at least 3 days, during which every The medium was changed twice a week to obtain primary hippocampal neurons. The method can be used to quickly and efficiently prepare primary hippocampal neurons, and the obtained primary hippocampal neurons are extremely suitable for use as cell models for patch clamp experiments.

Description

technical field [0001] The present invention relates to a method for culturing primary hippocampal neurons. Background technique [0002] Patch clamp technology is a new technology developed on the basis of voltage clamp technology, which has become the "gold standard" for studying ion channels. This technology is widely used in the study of ion channel gating kinetics, physiological functions, and pharmacological properties. and structure-function relationships. Primary rat hippocampal neurons are an important cell model for studying neuronal electrophysiological activity at the ex vivo level. Primary rat hippocampal neurons in good condition are an important prerequisite for successful patch clamp experiments, which are extremely important for sealing and membrane rupture in patch clamp recordings. [0003] At present, the culture methods of primary rat hippocampal neurons include serum culture and serum-free culture. The latest research is mostly based on serum-free cul...

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Problems solved by technology

[0003] At present, the culture methods of primary rat hippocampal neurons include serum culture and serum-free culture. The latest research is mostly based on serum-free culture methods, but the cell membrane of hippocampal neurons cultured without serum is relatively brittle. It is easy to deform after membrane rupture, which is not conducive to the recording of electrophysiological signals such as patch clamp, so it is not suitable for patch clamp experiments

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