In-vitro culture method of vaginal epithelial cells of mouse

Abstract

The invention relates to an in-vitro culture method of vaginal epithelial cells of a mouse and belongs to the technical field of cell culture. According to the method, isolated vaginal tissue of the mouse is sheared into pieces in the size of 1 cm*0.5 cm, washed and subjected to overnight culture through Dispase II, the vaginal tissue subjected to overnight culture is separated, lamina propria tissue is discarded, epithelial layer tissue is mixed with 0.25% pancreatin and 0.02% EDTA (ethylene diamine tetraacetic acid) and digested, finally, digestion is stopped through DMEM (Dulbecco modified Eagle medium) / F12 containing 10% fetal calf serum, obtained cell suspension is filtered, centrifugalized, re-suspended with a PBS (phosphate buffer solution) and EPILIFE sequentially and put into a 5% CO2 culture box with the temperature of 37 DEG C to be cultured, and a solution is replaced for the first time after two days and is replaced once every two days later. The method is convenient to operate, a large quantity of cells are obtained, the purity is high, the activity is high, the cells can be subcultured for 4-5 generations stably, and seed cells can be provided for observation of physiological and pathological change and the cancerization process as well as establishment of tissue-engineered vaginas of vaginal epithelium.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a method for culturing mouse vaginal epithelial cells in vitro, in particular to a method for primary culture of mouse vaginal epithelium by adopting an enzymatic digestion method. Background technique [0002] For the in vitro culture of mouse vaginal epithelial cells, domestic and foreign studies have used the tissue block method to culture vaginal epithelial cells. Although the operation is simple, it takes a long time and the passage cycle is long. [0003] In recent years, the mouse vaginal epithelium was isolated by enzymatic digestion, and the epithelial layer was digested into a single cell suspension by enzymatic digestion. After successful culture, it was used for tissue engineering vaginal research, but due to the However, due to the small number of cells obtained, the inability to adhere to the wall, and the poor ability of passage, there has not been...

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Problems solved by technology

[0002] For the in vitro culture of mouse vaginal epithelial cells, domestic and foreign studies have used the tissue block method to culture vaginal epithelial cells. Although the operation is simple, it takes a long time and the passage cycle is long

[0003] In recent years, the mouse vaginal epithelium was isolated by enzymatic digestion, and the epithelial layer was digested into a single cell suspension by enzymatic digestion. After successful culture, it was used for tissue engineering vaginal research, but due to the However, due to the small number of cells obtained, the inability to adhere to the wall, and the poor ability of passage, there has not been a large number of high-purity seed cells obtained in a relatively short period of time, which can be used to observe the physiological and pathological changes of the vaginal epithelium, the process of carcinogenesis, and organize Construction of the project

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