In vitro culture method of cervical intraepithelial neoplasia cells

Abstract

The invention discloses an in vitro culture method of cervical intraepithelial neoplasia cells. The in vitro culture method comprises the following steps: step one, taking a small piece of human cervical neoplasia tissue; step two, after digesting and decomposing by a type I collagenase, preparing a cell suspension; step three, utilizing a trypan blue dye exclusion method to judge the cell activity; step four, introducing the cell suspension into a culture bottle containing a low fetal bovine serum culture medium and coated with rat tail collagen, and waiting for the cells to adhere to the wall; and step five, after the cells adhere to the wall, replacing with a serum-free culture medium, purifying the cervical intraepithelial neoplasia cells, replacing the culture liquid weekly for at least two times, and after the cells are fused to 80%-85%, carrying out passage. The CIN cells cultured by the method have high survival rate and high purity, allows the morphology of the cells after passage to have no obvious change compared with that of the primary cells, and is carried with human papillomavirus (HPV); and the success of the CIN cell in vitro culture provides technical support for research on treatment of cervical cancer.

Description

technical field [0001] The invention relates to a cell culture method, in particular to a culture method for cervical intraepithelial neoplasia cells. Background technique [0002] Cervical cancer is the most common gynecological malignancy in the world. Its occurrence is a process that gradually evolves from precancerous lesions to cancer, and the time can range from several years to decades. This precancerous change represents a series of histologically abnormal cervical epithelial cells, so it is also called cervical intraepithelial neoplasia (CIN). According to the degree of cellular abnormalities, CIN is further divided into CINI (mild dysplasia), CINII (moderate dysplasia) and CINIII (severe dysplasia and carcinoma in situ). CINI will subside naturally in most cases, so in fact it does not belong to the category of precancerous lesions; however, CINII and CINIII often persist, and 20% to 45% of untreated CINII / III will further develop into cervical cancer, so , CINII...

AI Technical Summary

Problems solved by technology

The disadvantage of this culture method is that the culture method is relatively complicated, and the upper layer of the culture bottle needs to be cut off during the culture process, which increases the risk of contamination. At the same time, it takes a long time and needs five weeks for passage

Furthermore, FBS in the medium increases the risk of fibroblast contamination as well as differentiation of keratinocytes

Images