A kind of in vitro isolation and culture method and application of mammalian trigeminal ganglion cells

Abstract

The invention discloses an in vitro separation and culture method and application of mammalian trigeminal ganglion cells, belonging to the technical fields of cell biology and neurobiology. The method provided by the present invention is to place the mammalian trigeminal nerve tissue on a petri dish containing L-15 culture solution for cleaning treatment, and then cut it into pieces and then use type 11 collagenase, trypsin and trypsin-EDTA to digest it successively After the cell sieve is filtered, the filtrate is subjected to density gradient centrifugation, suspended and inoculated in a coated cell culture device, and cultured with a basic neural medium. The method of the present invention greatly reduces the damage to cells during tissue processing, removes a large number of cell fragments and other cell types and other impurities in the digestion process, greatly improves the purity of cells, and can obtain millions of relatively pure trigeminal nerve cells. The cell yield and quality are improved, and it is suitable for in vitro isolation and culture of mammalian trigeminal ganglion cells.

Description

technical field [0001] The invention relates to an in vitro separation and culture method and application of mammalian trigeminal ganglion cells, belonging to the technical fields of cell biology and neurobiology. Background technique [0002] Isolation and culture of neurons is a common technique for studying neuronal growth factors, axonal outgrowth, differentiation, and sensory physiology. Although it is relatively easy to isolate neurons from embryonic or neonatal animal neurons before they form synapses, embryonic neurons have different properties from mature neurons in terms of electrophysiology, development, and regeneration. Therefore, the culture of mature neurons is very necessary in many studies. However, in the process of primary neuron culture, there are always problems of low cell yield and quality. [0003] At present, the existing primary culture technology of trigeminal nerve cells is mostly the collagenase digestion method. During the culture process, som...

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Problems solved by technology

[0003] At present, the existing primary culture technology of trigeminal nerve cells is mostly the collagenase digestion method. During the culture process, some details are not considered comprehensively, such as: in the absence of CO 2 The stability of the culture medium is not good when the balance adjustment device is used, the treatment process has a greater impact on the tissue or cells, greater damage to the cells, insufficient digestion, more impurity cells, and impurity of the cultured cells, etc., so the yield of the cells and the quality is not high

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