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A method for constructing liver cell line of dark lip fish

A construction method and liver cell technology, applied in the field of freshwater aquatic organism cell culture, can solve the problems such as no similar reports, and achieve the effect of simple operation, large cell volume and good stability

Active Publication Date: 2018-10-26
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

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  • A method for constructing liver cell line of dark lip fish
  • A method for constructing liver cell line of dark lip fish
  • A method for constructing liver cell line of dark lip fish

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Prepare HBSS disinfection solution and cell culture solution

[0031] HBSS disinfectant: Add antibiotics to Hanks balanced salt solution to make the concentration of kanamycin sulfate 100μg / ml and the concentration of streptomycin sulfate 100μg / ml.

[0032] Basic culture medium: Add fetal bovine serum to the high-sugar DMEM culture medium so that the fetal bovine serum accounts for 10% of the total volume.

[0033] Primary culture medium: Add fetal bovine serum and antibiotics to the high glucose DMEM medium so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 150μg / ml and the concentration of streptomycin sulfate is 50μg / ml.

[0034] Subculture broth: Add fetal bovine serum and antibiotics to the high-sugar DMEM culture broth so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 50μg / ml and the concentration of streptomycin sulfate is 30μg / ml. .

[0035] Adjus...

Embodiment 2

[0045] 1. Prepare HBSS disinfection solution and cell culture solution

[0046] HBSS disinfectant: Add antibiotics to Hanks balanced salt solution to make the concentration of kanamycin sulfate 200μg / ml and the concentration of streptomycin sulfate 200μg / ml.

[0047] Basic culture medium: Add fetal bovine serum to the high-sugar DMEM culture medium so that the fetal bovine serum accounts for 10% of the total volume.

[0048] Primary culture medium: Add fetal bovine serum and antibiotics to the high-sugar DMEM medium so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 180μg / ml and the concentration of streptomycin sulfate is 100μg / ml.

[0049] Subculture broth: Add fetal bovine serum and antibiotics to the high-sugar DMEM culture broth so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 100μg / ml and the concentration of streptomycin sulfate is 50μg / ml. .

[0050] Adjus...

Embodiment 3

[0060] 1. Prepare HBSS disinfection solution and cell culture solution

[0061] HBSS disinfectant: Add antibiotics to Hanks balanced salt solution to make the concentration of kanamycin sulfate 300μg / ml and the concentration of streptomycin sulfate 300μg / ml.

[0062] Basic culture medium: Add fetal bovine serum to high-sugar DMEM culture medium so that fetal bovine serum accounts for 15% of the total volume.

[0063] Primary culture medium: Add fetal bovine serum and antibiotics to the high-sugar DMEM medium so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 200μg / ml and the concentration of streptomycin sulfate is 150μg / ml.

[0064] Subculture broth: Add fetal bovine serum and antibiotics to the high-sugar DMEM culture broth so that the fetal bovine serum accounts for 20% of the total volume. The concentration of kanamycin sulfate is 200μg / ml and the concentration of streptomycin sulfate is 150μg / ml. .

[0065] Adjust the p...

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Abstract

The invention relates to a method for establishing liver cell lines of semilabeo obscurus. The method includes steps of 1), acquiring the livers of the semilabeo obscurus: disinfecting the bodies of the semilabeo obscurus by methylene blue and alcohol jointly; 2), carrying out primary culture: carrying out culture on the livers by the aid of high-glucose DMEM (dulbecco modified eagle medium) culture fluid with kanamycin sulfate and streptomycin sulfate; 3), carrying out passage culture: replacing the cell culture fluid by basic culture fluid when the sixth generation of livers is subjected to passage culture, and successfully establishing the liver cell lines of the semilabeo obscurus. The method has the advantages that the obtained liver cell lines of the semilabeo obscurus are irregularly polygonal, can be subjected to continuous culture transfer and can be directly applied to research on biological characteristics, and requirements on conservation of resources of semilabeo obscurus species and theoretical research and application can be met.

Description

Technical field [0001] The invention relates to a method for establishing a cell line using dark-colored lip fish liver tissue, and belongs to the technical field of freshwater aquatic organism cell culture. Background technique [0002] The dark-colored lip fish Semilabeo obscurus Lin belongs to the Cyprinidae (Cyprinidae) genus Labeoninae (Semilabeo). Because the fish has fat body, tender meat and delicious taste, it has become the first choice for fishing. Because of its living habits in gregarious rock caves, it has become the biggest victim of illegal molecular electricity, poison, and explosion. With the increase in the use of fish resources and the updating of fishing technology, fish biodiversity is declining, with dark lips. Fish has become a rare and endangered species in the Lixianjiang River Basin. And the dark lip fish was listed as a second-level protected animal in Yunnan Province in 1989 and listed in the "Red Book of Endangered Animals in China" in 1998. In or...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12R1/91
Inventor 潘晓赋王晓爱杨君兴刘倩
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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