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In-vitro construction method and application of fugu rubripes ovarian cell line

A technology of red-finned pufferfish and ovarian cells, which can be applied to artificial cell constructs, animal cells, vertebrate cells, etc., can solve problems such as failure to establish, and achieve the effect of strong proliferation and passage ability and fast growth.

Inactive Publication Date: 2015-02-25
TIANJIN BOHAI SEA FISHERIES RES INST
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

So far, there are no relevant reports on the culture of puffer fish at home and abroad, and a cell line of puffer puffer has not been established to carry out toxicological research on the production of detoxified puffer fish and tetrodotoxin.

Method used

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  • In-vitro construction method and application of fugu rubripes ovarian cell line

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Embodiment Construction

[0050] The embodiments of the present invention will be described in further detail below in conjunction with the accompanying drawings. It should be emphasized that the following embodiments are illustrative, not restrictive, and should not be used as limitations of the present invention.

[0051] A method for constructing redfin puffer ovary cell line in vitro, the steps are as follows:

[0052] (1) Preparation of primary cell culture medium: the specific steps include:

[0053] ①Preparation of basal medium: 1000ml of MEM and L-15 medium mixed at 2:1 for basal medium (GIBCO standard), accurately weigh 2.38g of Hepes, dissolve in ultrapure deionized water, and mix evenly with magnetic stirring , adjust the pH at about 7.5, and then filter and sterilize, as a basic medium, it can be stored at 4°C for a long time (3 months);

[0054] ②Preparation of complete culture medium for puffer puffer ovary cells: 20% of the total volume of fetal bovine serum (FBS), 1% of turbot serum (S...

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Abstract

The invention relates to an in-vitro construction method and application of a fugu rubripes ovarian cell line. The construction method comprises the steps of (1) preparing a primary cell culture solution; (2) performing primary culture; (3) performing subculture; and (4) separating cell lines with different forms. The application comprises application of the fugu rubripes ovarian cell line to multiplication and expanding propagation of two typical fish viruses and application of the fugu rubripes ovarian cell line to transgenic research. According to the in-vitro construction method, an ovary is used as a culture and line construction object to build a biological platform for research on extraction and utilization of tetrodotoxin and safe detoxification treatment of cultured globefish, so that the conventional aquaculture industry and the emerging bio-pharmaceutical industry can be well served.

Description

technical field [0001] The invention belongs to the technical field of marine biological cell culture, in particular to an in vitro construction method and application of redfin puffer ovary cell lines. Background technique [0002] The red-finned oriental puffer belongs to the class of ray-finned fishes, the order Fuguformes, the family Tetraodontidae, and the genus Takifugu. "Puffer". As an important marine cultured fish in my country, pufferfish is not only delicious in meat, rich in protein and high in nutritional value, but also an important model fish. It is worth noting that the viscera, blood, ovaries and skin contain Tetrodonine, especially the ovary and liver. Therefore, the main raw materials for extracting Tetrodonine are the ovary and liver of the pufferfish. Tetrodotoxin (TTX) is an aminoperhydroquinazoline compound belonging to alkaloids, which was once considered to be the most toxic non-protein toxin in nature, and its toxicity is equivalent to 1250 times ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N7/00C12N5/10C12R1/91
Inventor 张博宋文平刘克奉郑德斌陈松林
Owner TIANJIN BOHAI SEA FISHERIES RES INST
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