In-vitro establishing method of acipenser ruthenus theca cell line and reagents applied to acipenser ruthenus theca cell line preparation

Abstract

The invention discloses an in-vitro establishing method of an acipenser ruthenus theca cell line and reagents applied to acipenser ruthenus theca cell line preparation. The in-vitro establishing method of the acipenser ruthenus theca cell line includes the steps: 1, a digestive juice is used for digesting acipenser ruthenus ovarian tissue to obtain an acipenser ruthenus ovarian tissue cell suspension; 2, a standing-centrifugation method is adopted to obtain acipenser ruthenus theca cells, wherein the acipenser ruthenus ovarian tissue cell suspension is subjected to standing, a supernatant containing cells and cell masses is collected and then centrifuged, a sediment is collected, a culture solution is added into the sediment to prepare a cell suspension, and inoculation and culture are conducted to obtain the acipenser ruthenus theca cells; and 3, digestion is conducted two times, and subculture is conducted, wherein after the acipenser ruthenus theca cells grow into a single layer, the digestive juice is adopted to carry out digestion two times, and then subculture is carried out to obtain the acipenser ruthenus theca cell line. According to the in-vitro establishing method of theacipenser ruthenus theca cell line, the standing-centrifugation method and a two-time digestion method are adopted to establish the acipenser ruthenus theca cell line, the established acipenser ruthenus theca cell line can be continuously subcultured, the cells obtained through the in-vitro establishing method can be subcultured for 60 passages or above, and a large number of theca cells can be provided in vitro.

Description

technical field [0001] The invention belongs to the technical field of fish cell culture, and in particular relates to an in vitro construction method of an ancient sturgeon fish Acipenser ruthenus (Acipenser ruthenus) follicular theca cell line and a reagent used therefor. Background technique [0002] Sturgeon is an ancient, economically important, large-scale sub-cold water fish that originated 200 million years ago and is known as a "living fossil in water". There are currently 27 species of sturgeon in the world. All sturgeons are currently listed as second-class protected animals in the appendix of the International Convention on the Protection of Endangered Species of Animals and Flora. At the same time, due to its delicious meat, caviar has the reputation of "egg gold", and the market demand for sturgeon roe, fish meat and other products is increasing year by year. With the continuous improvement of artificial reproduction technology, the research team has achieved...

AI Technical Summary

Problems solved by technology

The traditional cell culture method is not suitable for the establishment of the theca cell line of sterlet sturgeon, the number of cells obtained by the traditional method is small, the cell activity is poor, and the primary culture cannot be initiated or passaged

The in vitro culture of fish ovarian tissue cells has always been a difficult point in fish cell culture. So far, no report has been made on the theca cell line of sturgeon

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