Method for separating and culturing trigeminal root ganglion neurons of mammal in vitro and application of method

Abstract

The invention discloses a method for separating and culturing trigeminal root ganglion neurons of a mammal in vitro and application of the method, and belongs to the technical field of cell biology and neurobiology. The method comprises the following steps: placing the trigeminal root ganglion neurons of the mammal in a culture dish containing L-15 culture solution for washing treatment, cutting the trigeminal root ganglion neurons into pieces and then carrying out digestion treatment with type 11 collagenase, trypsin, and trypsin-EDTA in sequence, filtering with a cell strainer and then carrying out density gradient centrifugation for the filtrate, suspending and inoculating the obtained product to a wrapped cell culture device, and culturing with a basic neuronal medium. The method greatly reduces the damage to the neurons in the tissue processing process, removes the impurity contamination in the digestion process like cell debris and other cells, greatly improves the purity of the neurons, can obtain a million of pure trigeminal ganglion neurons, improves the yield and the quality of the trigeminal ganglion neurons, and is suitable for the in-vitro separation and culturing of the trigeminal root ganglion neurons of the mammal.

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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