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Construction method of sinocyclocheilus grahami grahami fin cell line

A construction method and technology of fin cells, which are applied in the field of freshwater aquatic organism cell culture, can solve problems such as no identical reports of inventions, meet the needs of germplasm resource preservation, theoretical research and application, and are easy to operate

Inactive Publication Date: 2012-10-31
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

[0004] Through document retrieval, do not see identical report with the present invention

Method used

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Experimental program
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Embodiment Construction

[0020] 1. Preparation of cell culture medium

[0021] Select the L-15 medium with a pH value of 7.2-7.4, add fetal bovine serum and human basic fibroblast growth factor to the medium, so that the final concentration of fetal bovine serum is 20%, and human basic fibroblasts grow The final concentration of the factor is 5 ng / ml, and it is stored in a refrigerator at 4°C for later use;

[0022] 2. Minimally invasive sampling of fin tissue

[0023] Disinfect the live fish in a solution containing 8 mg / L potassium permanganate for 10 min, then remove the caudal fin with a sterile instrument, place it in a 12-well plate, add 200 IU / ml penicillin, 200 μg / ml streptomycin and 15 μg / ml amphotericin B in HBSS solution 3 ml. After disinfection, the fish body was put back into the fish tank for rearing. After 3 months of daily care, fin rays re-grow;

[0024] 3. Primary culture

[0025] Remove the caudal fin tissue in a sterile operating bench, wash it three times with a solution of HBS...

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Abstract

The invention relates to a construction method of a sinocyclocheilus grahami grahami fin cell line and belongs to the technical field of freshwater aquatic organism cell culture. The method comprises that a sinocyclocheilus grahami grahami fin ray tissue serves as a material, a tissue block method is adopted, and culturing is achieved in a L-15 culture solution containing fetal calf serum and human basic fibroblast growth factors and having a PH value ranging between 7.2 and 7.4; subculture is performed by using a trypsin digestion method; and the subculture particularly comprises preparation of cell culture fluid, fin tissue minimally invasive sampling, primary culture and continuous culture. The method has the advantages that 1, the minimally invasive processing is adopted, on the premise that the fish is alive, and the operation is easy; 2, the sinocyclocheilus grahami grahami fin cell line is in a fibroblast shape, continuous passage can be achieved, the line can be applied to biological characteristic research directly, and requirements for sinocyclocheilus grahami grahami germplasm conservation and theoretical research and application are met; and 3, the construction method is also applicable to cell lines of constructed fin rays of other fishes.

Description

Technical field: [0001] The invention relates to a method for constructing a cell line of golden-threaded barb fin in Dianchi Lake, and belongs to the technical field of freshwater aquatic organism cell culture. Background technique: [0002] The golden-threaded barb Sinocyclocheilus grahami (Regan, 1904) belongs to the Cypriniformes (Cypriniformes) Cyprinidae (Cyprinidae) genus Sinocyclocheilus, and is endemic to the Dianchi Lake Basin. In 1989, the golden-threaded barb in Dianchi Lake was listed as a national second-class protected animal, listed as an endangered level in the "China Red Book of Endangered Animals (Fish)", and listed as a VU level in the Chinese Red List of Species. The fish is named for its golden side line scales shining in the sun, like golden threads. The fish is tender and delicious, and it ranks first among the four famous fish in Yunnan. [0003] Since the late 1950s, due to the invasion of alien species, water pollution, habitat loss, overfishing a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12R1/91
Inventor 杨君兴王晓爱潘晓赋陈小勇
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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