Fish embryo cell separation and culturing method

Abstract

The separation and cultivation method for fish embryonic cell comprises: collecting cell; in CM, separating the cell of blastula-archenteron late time with laceration method, and culturing the initial and passage cells; identifying the cell on morphology, proliferation feature and chromosome karyotype. Wherein, using 70% alcohol to dip and sterilize for embryo; culturing cell in common incubator with 24Deg to passage 2-3 times every week. The objective cell is litle, circle or polygon, and growing stably, and has high rate of diploid cell. This invention is simple and convenient, and can be applied into separation and cultivation for almost all fish embryonic cell.

Description

Technical field: [0001] The invention belongs to the culturing technology of fish cells in the aquatic product technology, and is a technical method for obtaining an embryonic cell line with infinite proliferation ability by using fish embryo cells as materials under artificial culture conditions in vitro. Background technique: [0002] Fish embryo cells are cells with immortal proliferation capacity isolated and cultured from fish blastocyst stage to late gastrulation stage embryos. Before the present invention was made, there were no relevant reports on the separation and culture of embryonic cells of marine aquaculture fish at home and abroad. In terms of embryo-derived cell culture, the international community mainly focuses on the research of small-scale fish-zebrafish and medaka embryo cell culture: Collodi et al. (2004) conducted research on the isolation and culture of zebrafish embryo cells. It is a trophoblast cell culture method, that is, a single layer of rainbow trou...

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Problems solved by technology

In terms of embryo-derived cell culture, the international research mainly focuses on the cell culture of small model fish—zebrafish and medaka embryos: Collodi et al. (2004) carried out research on the isolation and culture of zebrafish embryo cells. It is a trophoblast cell culture method, that is, a single layer of rainbow trout spleen cells (RTS34st) is first cultured, and then cells isolated from embryos are placed on a single layer of rainbow trout spleen cells for culture. After the zebrafish embryonic cell colony grows, Transfer the colony to a test tube with a Pasteur glass pipette, digest it into a single cell, form a suspension, and place the cell suspension on a single layer of rainbow trout splenocytes for subculture; Lannan CN et al. (1984) established the Damaha Fish embryonic cell lines, the embryonic cell separation method they use is complicated and not easy to succeed; the cell culture process is cumbersome, and the cell growth rate is slow; it is difficult to cultivate a large number of cells, which is not conducive to using cells to do a large number of experiments

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