Separating and culturing method for olfactory ensheathing cells of rats

Abstract

The invention provides a separating and culturing method for olfactory ensheathing cells of rats. The separating and culturing method comprises the following steps: (1) disinfecting a tissue processing instrument; (2) selecting male SD rats of 2-3 months old, killing the male SD rats after anesthesia, separating the olfactory mucosa tissue by adopting surgical scissors, placing the tissue in PBSsolution which is precooled, is sterile and contains double antibodies, and washing the tissue for a plurality of times, so that blood stains and outer membranes are removed; (3) cutting up the tissue, and carrying out digestion by adopting preheated neutral enzyme and trypsin respectively; (4) terminating the digestion, carrying out centrifugation, and discarding the supernatant; (5) inoculatingthe obtained product into an enveloped culture flask by adopting an artificial wall sticking method; (6) carrying out incubation for 1-2 h, and carrying out culturing in the environment being at 37 DEG C containing 5% CO2; (7) carrying out cell purification; (8) carrying out cell morphological observation; (9) carrying out cell subculturing; (10) carrying out cell counting and survival rate testing; and (11) carrying out immunological identification. For the separating and culturing method for olfactory ensheathing cells of rats, the operation is simple, a large number of cells are obtained, the survival rate is high, the method belong to an ideal primary separating and culturing method for the olfactory ensheathing cells of rats, and a reliable cell resource is provided for the experiment.

Description

technical field [0001] The invention belongs to the technical field of cell culture of modern biotechnology, and in particular relates to a method for separating and culturing rat olfactory ensheathing cells. Background technique [0002] The olfactory mucosa is a pseudostratified columnar epithelium that contains a variety of cellular components, mainly including olfactory neurons and supporting cells. There are fibroblasts, epithelial cells, and olfactory ensheathing cells in the olfactory mucosa. Olfactory ensheathing cells (OECs) are a unique type of glial cells in the central nervous system, distributed in the olfactory bulb and olfactory mucosa of the nasal cavity. Functionally between Schwann cells and oligodendrocytes. OECs can inhibit glial growth, neurotrophy, scar formation, and sheath formation. It provides a suitable microenvironment for the growth of axons, and is one of the ideal candidate cells for promoting the regeneration of the central nervous system. ...

AI Technical Summary

Problems solved by technology

[0005] At present, most of the isolation and culture methods of rat olfactory ensheathing cells are based on enzymatic digestion, and the purity of the isolated cells is not high, and the culture time is long

Regarding the cultivation of olfactory ensheathing cells, the methods introduced at home and abroad have problems such as poor reproducibility or complexity, which is not conducive to practical application

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