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In-vitro culture method of triploid loach fin cells

An in vitro culture and cell culture technology, applied to artificial cell constructs, animal cells, vertebrate cells, etc., can solve the problems of easy floating of cells, difficult growth, microbial contamination, etc., to overcome microbial contamination, simple steps, and easy operation Effect

Inactive Publication Date: 2013-02-13
DALIAN OCEAN UNIV
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AI Technical Summary

Problems solved by technology

However, due to problems such as microbial contamination, easy floating of cells, and difficulty in in vitro growth in fish fin cell culture, there has been no relevant report on the in vitro culture of triploid loach fin cells.

Method used

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  • In-vitro culture method of triploid loach fin cells
  • In-vitro culture method of triploid loach fin cells

Examples

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Embodiment 1

[0015] a. Preparation of cell culture medium

[0016] Select the DMEM / F12 medium of Hyclone Company, add fetal calf serum, human basic fibroblast growth factor, type I insulin-like growth factor, epidermal growth factor and chondroitin sulfate to the medium to prepare cell culture medium, wherein Fetal bovine serum accounts for 20% of the total volume, the concentration of human basic fibroblast growth factor is 10ng / ml, the concentration of type I insulin-like growth factor is 20ng / ml, the concentration of epidermal growth factor is 10ng / ml, cartilage sulfate The concentration of the element is 10ug / ml; it is stored at 4°C;

[0017] b. Primary culture

[0018] After the loach is temporarily raised in high-double-antibody disinfection aeration water (1000IU / mL penicillin, 1000μg / mL streptomycin) for 16~24h, it is treated with 1‰ of benzene containing high double-antibody (1000IU / mL penicillin, 1000μg / mL streptomycin). After methanol anesthesia, take the fin tissue of triploi...

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Abstract

The invention discloses an in-vitro culture method of triploid loach fin cells. The method includes selecting DMEM / F12 culture medium, and adding fetal bovine serum (FBS), human basic fibroblast growth factor (hbFGF), type-I insulin-like growth factor, epidermal growth factor (EGF) and chondroitin sulfate to prepare cell culture solution; rinsing triploid loach fin with povidone-iodine (PVP-I) and penicillin-streptomycin on a working bench; performing primary culture in a cell culture bottle until a single layer is formed; preparing into a cell suspension; adding culture medium, and performing subculture. The procedure is easy and simple in operation. The cultured triploid loach fin cell is in fibrous morphology, can be sub-cultured successively and directly applied to multiploid fish biological research, virus separation and proliferation, vaccine development and functional gene research.

Description

technical field [0001] The invention relates to an in vitro culture method for loach fin cells, in particular to an in vitro culture method for triploid loach fin cells that is simple to operate and can be used for polyploid fish biological research, virus isolation and amplification, vaccine development and functional gene research method. Background technique [0002] loach( Misgurnus anguillicaudatus) In taxonomy, it belongs to Cypriniformes, Cobitidae, Cobitinae, and Misgurnus. It is a wild small freshwater fish unique to Asia, widely distributed in most of the south of the Liaohe River in my country. region as well as Vietnam, Korea and Japan. It is distributed in various freshwater waters in my country, such as lakes, ponds, rivers, ditches, and rice fields. Loach meat is tender, delicious, and rich in nutrition. It is known as "ginseng in water". In addition to its high edible value, it also has certain nourishing and medicinal effects, and is one of the best-sellin...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 李霞马辰李雅娟秦艳杰
Owner DALIAN OCEAN UNIV
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