Method for culturing primary hippocampal neurons

Abstract

The invention discloses a method for culturing primary hippocampal neurons, wherein the method comprises the following steps: (1) digesting a hippocampal tissue; (2) blowing and dispersing the digested hippocampal tissue so as to obtain a cell suspension; (3) inoculating the cell suspension and culturing the hippocampal tissue for the first time for 2-4h so as to promote cell attachment; (4) changing a first culture solution inside a culture dish with a second culture solution, and culturing for the second time for 24h; (5) adding 1-2ug / ml of cytarabine to the culture dish and culturing for the third time for 24h; and (6) changing half volume of solution inside the culture dish by virtue of the second culture solution, culturing for the fourth time for at least 3 days and meanwhile changing half volume of solution twice every week so as to obtain the primary hippocampal neurons. By virtue of the method, primary hippocampal neurons can be rapidly and efficiently prepared and acquired; and the acquired primary hippocampal neurons are quite suitable to act as a cell model in a patch clamp experiment.

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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