Mechanical separation method of neurons of adult mice

Abstract

The invention which relates to the biomedicine field discloses a mechanical separation method of neurons of adult mice. The method comprises the following steps: making the adult mice's brain tissue slices with the thickness of 350-375mum in an ice bath tissue culture solution with an electronic vibration slicer, preserving for 30-60min at room temperature, putting the brain tissue slices into a papain-containing tissue culture solution under aseptic conditions, controlling the temperature at 32.2-35.3DEG C, maintaining the temperature for 30min, placing the brain slices in a cell culture dish containing an HEPES buffer solution, selecting a neural region needing separation, and separating neurons through using a cell separator; and screening healthy neurons. The method of the invention, which allows a primary culture chemical separation method to be integrated into mechanical cell separation and the brain slices to be preprocessed with the papain, makes the healthy neurons of adult and aged mice to be obtained through the mechanical separation, and has the advantages of simplicity, convenience, and substantial separation effect.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for mechanically separating adult mouse neurons. Background technique [0002] The patch clamp experiment of mechanically isolated neurons has an irreplaceable role in the study of neurotransmitter receptor mechanisms. Mechanically isolated neurons have better physiological channel mechanisms than primary cultured neurons, and are usually used to Most of the mice subjected to mechanical separation experiments were 2-3 weeks old (young). The brain structure of old mice is dense, and it is basically impossible to perform mechanical separation, which brings difficulties to the comparison of neurotransmitter receptor mechanisms in mice of different ages. Contents of the invention [0003] The purpose of the present invention is to provide a method that integrates the chemical separation method of primary culture into mechanical cell separation, treats brain slices with papain in...

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Problems solved by technology

[0002] The patch clamp experiment of mechanically isolated neurons has an irreplaceable role in the study of neurotransmitter receptor mechanisms. Mechanically isolated neurons have better physiological channel mechanisms than primary cultured neurons, and are usually used to Most of the mice subjected to mechanical separation experiments were 2-3 weeks old (young). The brain structure of old mice is dense, and mechanical separation is basically impossible, which makes it difficult to compare the mechanism of neurotransmitter receptors in mice of different ages.

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