Immortalized goat small intestine epithelial cell line and establishment method thereof

Abstract

The present invention provides an immortalized goat small intestine epithelial cell line and an establishment method thereof. The method comprises: culturing primary goat small intestine epithelial cells through tissue block, immortalizing the primary-culture goat small intestine epithelial cells, purifying the goat small intestine epithelial cells through a labelling scraping method, and finally establishing the immortalized goat small intestine epithelial cell line. According to the present invention, the addition of any cell growth factors during the immortalized goat small intestine epithelial cell culture process is not required, the operation is convenient and simple, and a lot of cost is saved compared to the primary cell culture; and the method sequentially comprising purifying and cell immortalizing is generally used in the prior art while the labeling method is used to gradually scrap and purify the immortalized goat small intestine epithelial cells, such that the defect that the aging of the goat small intestine epithelial cells due to the prolonged culture time finally causes the cell immortalization failure in the prior art can be avoided.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to an immortalized goat small intestinal epithelial cell line and a method for establishing the same. Background technique [0002] Goat small intestinal epithelial cells are of great significance to intestinal nutrient absorption and metabolism, the relationship between microbial flora and as an immune barrier in goat intestinal tract. Foreign scholars have done a lot of research on small intestinal epithelial cells and intestinal immune mechanisms, mainly using human, mouse and rat small intestinal epithelial cells as models [1-2]. At present, there are few studies on the primary culture of small intestinal epithelial cells from ruminants. In the method of obtaining primary small intestinal epithelial cells through culture, the enzyme digestion method is mainly used. During the process of digesting tissue blocks, enzyme digestion will irreversibly damage the receptors on the surfa...

AI Technical Summary

Problems solved by technology

Therefore, the primary goat small intestinal epithelial cells cultured in vitro can only be subcultured for 3-5 passages, after which the cells cannot proliferate and undergo apoptosis. For example, Kuang Wei, a researcher in our laboratory, obtained goat small intestinal epithelial cells in 2007. When passed to the third generation, a large number of cells are senescent and apoptotic, unable to proliferate, and ultimately make subsequent experiments impossible[8]

[0005] In summary, there are the following technical difficulties in the immortalization of goat small intestinal epithelial cells based on tissue block culture: 1) If monoclonal purification of goat small intestinal epithelial cells is performed first, it will lead to apoptosis and cannot Perform subsequent cell immortalization operations; 2) If the unpurified goat intestinal epithelial cells are directly subjected to cell immortalization operations, then fibroblasts will also be immortalized, because the proliferation rate of immortalized fibroblasts is much greater than that of epithelial cells cells, then the fibroblasts growing together with the epithelial cells will surround the goat small intestinal epithelial cells, which will cause the epithelial cells to be squeezed by the fibroblasts, and eventually the immortalized goat small intestinal epithelial cells will fall off the bottom of the culture flask

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