In-vitro separation culture method for hippocampal neurons of adult rat

Abstract

The invention discloses an in-vitro separation culture method for hippocampal neurons of an adult rat. The in-vitro separation culture method comprises the following steps of coating a culture plate; separating a rat brain; separating a hippocampus; preparing a cell suspension; further purifying the neurons; carrying out cell inoculation; and carrying out in-vitro primary culture. The invention aims at providing a high-purity and high-activity acquiring (separating and purifying) method for hippocampal neurons and an establishing method for a flow culture system suitable for long-term in-vitro culture of hippocampal neurons of the adult rat.

Description

technical field [0001] The invention belongs to the technical field of life science / neurobiology, and specifically relates to a scheme for isolating and culturing adult rat hippocampal neurons in vitro. Background technique [0002] Neurons are the basic units that constitute the structure and function of the nervous system. The primary culture of rat brain neurons in vitro has been widely used as a model for studying the physiology and pathology of the central nervous system. However, in the current research, most reports focus on the culture of fetal mouse or neonatal mouse neurons. Neurons from adult mice are difficult to culture in vitro, so there are few reports. [0003] Previous studies have shown that embryonic / neonatal neurons behave differently from adult neurons in many aspects such as pharmacology, electrophysiology, development, regeneration, and pathological characteristics. Certain age-related diseases such as Alzheimer's disease and Parkinson's disease lim...

AI Technical Summary

Problems solved by technology

Frequent medium changes lead to a large loss of neurotrophic factors secreted by neuronal metabolism into the culture medium, which affects the long-term culture of neurons in vitro

[0008] The following four problems generally exist in the existing primary culture models of neurons in vitro: (1) low purity

The cultured neurons are often mixed with other cell types such as glial cells; (2) Poor activity

During the separation process of adult neurons, the tight junctions between axons, dendrites, neuron cell bodies, and the synaptic structure formed by the cross-linking of different neurons are severely damaged, and regeneration in vitro is difficult; (3) Long-term culture is not possible

The existing artificially set in vitro culture environment cannot meet the needs of long-term culture of neurons; (4) Static model

It is impossible to truly simulate the physiological conditions in the body. The renewal of nutrients and the elimination of metabolic wastes mainly rely on the diffusion movement of molecules and frequent fluid changes.

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