Pseudosciaena crocea head kidney cell line and construction method thereof

A large yellow croaker and kidney cell technology, applied in the field of cell lines, can solve the problems of increased mortality of large yellow croaker and affecting the healthy development of large yellow croaker aquaculture

Active Publication Date: 2015-07-01
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In recent years, with the increasing environmental pollution and the overexpansion of the aquaculture industry, the problem of large yellow croaker diseases has become increasingly serious, and various infectious diseases have frequently broken out, causing the mortality rate of large yellow croakers to increase year by year and causing serious economic losses, which greatly affected the large yellow croaker aquaculture industry. healthy development of

Method used

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  • Pseudosciaena crocea head kidney cell line and construction method thereof
  • Pseudosciaena crocea head kidney cell line and construction method thereof
  • Pseudosciaena crocea head kidney cell line and construction method thereof

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Embodiment Construction

[0022] One, the construction method of large yellow croaker head kidney tissue cell line, the steps are as follows:

[0023] 1. Preparation of cell culture medium: Take a bag of L-15 medium powder from GIBCO Company, add sterile deionized water to dissolve, adjust the pH to 7.6-7.7 with an appropriate amount of 3.6% hydrochloric acid, and then add 2.5mL sterile HEPES buffer Add sterile deionized water to 900mL, filter with 0.22μm filter membrane, subpackage, take a small amount for sterility test, store at 4°C, and use it as the basic medium; add appropriate amount of fetal bovine serum and double Anti-(1× double antibody is penicillin 100U / mL, streptomycin 0.1mg / mL), stored at 4°C, it is a complete medium.

[0024] 2. Establishment of primary culture: Take 3 healthy large yellow croakers in the sterile room, measure their body length, and weigh them. Immerse in 75% ethanol, perform body surface disinfection and anesthesia, and dissect immediately. Prepare a sterile glass bo...

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Abstract

The invention discloses a pseudosciaena crocea head kidney cell line and a construction method thereof, relating to cell lines. The pseudosciaena crocea head kidney cell line is a pseudosciaena crocea head kidney cell line LYCK with the preservation number of CCTCC NO: C2013113. The construction method of the pseudosciaena crocea head kidney cell line comprises the steps of (1) preparing a cell culture medium; (2) constructing primary culture; (3) performing secondary culture. The constructed pseudosciaena crocea head kidney cell line is fibrous, can be subjected to continuous passage, and is sub-cultured for 70 generations; a large quantity of pseudosciaena crocea head kidney cell lines can be provided; the pseudosciaena crocea head kidney cell line can be directly applied to research of various pseudosciaena crocea immunity related functional genes. The pseudosciaena crocea head kidney cell line can be applied to important researches including cytobiology, molecular biology, virology, toxicology, oncology, genetics, immunology and the like.

Description

technical field [0001] The invention relates to a cell line, in particular to a large yellow croaker head kidney cell line and a construction method thereof. Background technique [0002] At present, there are at least thousands of cell strains and cell lines established in the world, including normal cells, tumor cells, and genetically defective cells of humans and animals. Among them, the cell culture of marine organisms is later than that of terrestrial organisms. The rapid development of the fish farming industry has also promoted the establishment of fish cell lines and their applications in virus detection, vaccines and pathotoxicology. The establishment of fish cell lines began in the 1960s. In 1962, Wolf et al. (WOLF K, QUIMBY M.Established eurythermic line of fish cells in vitro.Science1962.135:1065-1066.) established the rainbow trout gonad cell line RTG-2 for the first time, which has been subcultured in the laboratory so far . Since then, the culture of fish c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12R1/91
Inventor 陈新华敖敬群王贤慧王琨茹
Owner THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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