A method for constructing muscle cell line of soft-finned lobster fish

Abstract

The invention relates to a method for building a neolissochilus banasi muscle cell line. The method comprises the following steps: (1) neolissochilus banasi muscle acquisition: using potassium permanganate and alcohol together to disinfect a fish body; (2) primary culture: using a DMEM culture solution containing kanamycin sulfate, streptomycin sulfate and amphotericin B for culture; (3) subculture: at the 10th generation, replacing a cell culture solution with a basal culture solution and successfully building the neolissochilus banasi muscle cell line. By adoption of the method, the acquired neolissochilus banasi muscle cell line takes the shape of an irregular polygon, and can realize continuous cell culture and can be directly applied to researches on bionomics, thereby meeting the demands for germplasm resource conservation and theoretical researches, as well as application, of neolissochilus banasi.

Description

technical field [0001] The invention relates to a method for establishing a cell line by using the muscle tissue of the soft-finned lip fish, and belongs to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. Background technique [0002] Neolissochilus benasi ((Pellegrin&Chevey, 1936)), formerly known as soft-finned barb, belongs to the genus Neolissochilus (Cypriniformes) Cypriniformes (Cypriniformes) subfamily (Barbinae) Neolissochilus, mainly Distributed in the Red River system. With the development of hydropower in the Red River system in China, it has had many adverse effects on a variety of rare indigenous fish in the river. Therefore, in 2007, the Yunnan Provincial Environmental Protection Bureau approved the project, and Yunnan Datang International Lixian River Basin Hydropower Development Co., Ltd. entrusted China The Kunming Institute of Zoology of the Academy of Sciences carried out research on the artifici...

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Problems solved by technology

[0004] Although the artificial propagation technology of the soft finned lobster has been successfully developed, and it has been released back to the Lixianjiang River Basin in a planned way, due to the limitations of the cultured fish itself, artificial reproduction in the pond culture environment is likely to cause chromosomal abnormalities, such as polyploidy and aneuploidy It requires short-term cell culture for karyotype analysis to test the quality of artificially propagated species and ensure the survival rate and reproduction rate of released individuals

However, for the larvae of soft-finned eucalyptus, which grow slowly, are relatively small, and have low blood volume, the traditional short-term blood culture cannot meet the needs of karyotype analysis. Therefore, it is necessary to find a method that can quickly obtain cells. A Method to Meet the Short-Term Culture of Softfin Neophytes Cells for Karyotyping

[0005] Fish cell culture started in the 1960s, and more than 280 cell lines have been established so far. After more than 30 years of fish cell culture in China, only more than 50 cell lines have been established. The species of cell lines established are not enough for Chinese fish 1.5% of the total (more than 3,500 species of fish in China), it can be seen that the construction speed of cell lines is very slow, and insufficient attention and fewer participating workers may be one of the reasons. In addition, compared with breastfeeding Animal cell culture and fish cell culture are more likely to be polluted, leading to an increase in failure rate, but most of the existing patents or articles attribute this phenomenon to poor technology, and pay too little attention to the quality of the fish itself. In the practice of cell culture, the living conditions of the fish directly affect the success or failure of the cell line construction. Therefore, the successful construction of the muscle cell line of the soft-finned fish is based on the wild and pond culture of the soft-finned fish. Based on an excellent understanding of ecological habits

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