Construction method for normal and whitening skin cell line of bastard halibut

Abstract

The invention belongs to the technical field of marine organism cell culture and specifically relates to a construction method for a normal and whitening skin cell line of bastard halibut. The method comprises the following steps: firstly preparing a cell culture fluid, taking normal and whitening skin tissues of the bastard halibut as materials, rinsing and cutting, then directly putting into a 25cm<2> culture flask filled with a defined amount of cell culture fluid, and culturing at 24 DEG C; exchanging the cell culture fluid every 5 days, and adopting 0.25% trypsin for digesting cells for sub-culturing when the original generation cell grows into a single layer; and sub-culturing for one time per 8-10 days, and successfully constructing a cell line when the tenth generation is sub-cultured and the serum content in the cell culture fluid is reduced from 20% to 10%. The cell line of bastard halibut, acquired according to the method, is in a fiber form, more than 40 generations can be continuously sub-cultured, and the cell line can be directly applied to the etiologic characteristic research, vaccine research and functional gene research. The construction method is suitable for constructing the skin cell lines of the other fishes.

Description

technical field [0001] The invention belongs to the technical field of marine biological cell culture, and in particular relates to a method for constructing normal and albino skin cell lines of flounder. Background technique [0002] Since the first teleost cell line—the rainbow trout gonad cell line RTG-2 (Wolf and Quimby, Science, 1962, 135:1065-1066) was successfully constructed in 1962, nearly 300 different fish cell lines have been developed successively Established, tissue sources include embryonic and adult fish kidneys, hearts, spleens, fin rays, gonads, etc. The application of these cell lines has also been extended from virus isolation and identification to research in the fields of fish embryonic stem cells, immunology, etiology, genetic breeding and genetic engineering, and fish resource protection. However, fish skin is an important pathogen defense barrier and a tissue for pigmentation. So far, only a few fish have skin cell lines (Lei et al., Fish Physiology...

AI Technical Summary

Problems solved by technology

This bleaching has seriously affected the market value and individual viability of flounder fry

So far, the research on the albino mechanism of flounder only stays in the aspects of bait nutrition, environmental conditions and physiological factors, and there are few reports on the molecular mechanism

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