Method for establishing liver cell lines of semilabeo obscurus

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

Active Publication Date: 2015-08-05
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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  • Method for establishing liver cell lines of semilabeo obscurus
  • Method for establishing liver cell lines of semilabeo obscurus
  • Method for establishing liver cell lines of semilabeo obscurus

Examples

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Effect test

Embodiment 1

[0030] 1. Preparation of HBSS disinfection solution and cell culture solution

[0031] HBSS disinfectant: add antibiotics to Hanks balanced salt solution, so that the concentration of kanamycin sulfate is 100 μg / ml, and the concentration of streptomycin sulfate is 100 μg / ml.

[0032] Basal culture medium: add fetal bovine serum to the high-sugar DMEM 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-sugar DMEM medium, so that the fetal bovine serum accounts for 20% of the total volume, the concentration of kanamycin sulfate is 150 μg / ml, and the concentration of streptomycin sulfate is 50 μg / ml ml.

[0034] Subculture 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 50 μg / ml, and the concentration of streptomycin sulfate is 30 μg...

Embodiment 2

[0045] 1. Preparation of HBSS disinfection solution and cell culture solution

[0046] HBSS disinfectant: add antibiotics to Hanks balanced salt solution, so that the concentration of kanamycin sulfate is 200 μg / ml, and the concentration of streptomycin sulfate is 200 μg / ml.

[0047] Basal culture medium: add fetal bovine serum to the high-sugar DMEM 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 180 μg / ml, and the concentration of streptomycin sulfate is 100 μg / ml ml.

[0049] Subculture 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 100 μg / ml, and the concentration of streptomycin sulfate is 50 ...

Embodiment 3

[0060] 1. Preparation of HBSS disinfection solution and cell culture solution

[0061] HBSS disinfectant: add antibiotics to Hanks balanced salt solution, so that the concentration of kanamycin sulfate is 300 μg / ml, and the concentration of streptomycin sulfate is 300 μg / ml.

[0062] Basal culture medium: add fetal bovine serum to the high-sugar DMEM medium, so that the 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 200 μg / ml, and the concentration of streptomycin sulfate is 150 μg / ml ml.

[0064] Subculture 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 200 μg / ml, and the concentration of streptomycin sulfate is 150...

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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 by using the liver tissue of the dark lip fish, and belongs to the technical field of freshwater aquatic organism cell culture. Background technique [0002] Semilabeo obscurus Lin belongs to the genus Semilabeo of the subfamily Labeoninae of Cyprinidae. Because of its fat body, tender meat and delicious taste, it has become the first choice for fishing. And because of its living habit of living in caves in groups, it has become the biggest victim of electricity, poison, and bombing by criminals. With the increase in the utilization of fish resources and the update of fishing technology, the biodiversity of fish is declining. Dark lips Fish has become a rare and endangered species in the Lixianjiang River Basin. In 1989, the dark-colored lip fish was listed as a Class II protected animal in Yunnan Province, and in 1998 it was included in the "China Red Data Book of Endangered Animals". I...

Claims

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Application Information

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Patent Type & Authority Applications(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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