A method for constructing a rhino-horned catfish heart cell line

A rhinoceros horn catfish and construction method technology, applied in artificial cell constructs, microbial-based methods, animal cells, etc., can solve problems such as no similar reports, achieve repeatability, low pollution rate, and simple and easy operation line effect

Active Publication Date: 2019-03-08
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 used

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  • A method for constructing a rhino-horned catfish heart cell line
  • A method for constructing a rhino-horned catfish heart cell line
  • A method for constructing a rhino-horned catfish heart cell line

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1. Prepare PBS disinfectant and cell culture medium

[0027] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 100IU / ml, the concentration of streptomycin is 100μg / ml, the concentration of streptomycin sulfate is 40μg / ml, and the concentration of amphotericin B is 40μg / ml.

[0028] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 300 μg / ml, and the fetal bovine serum accounts for 15% of the total volume.

[0029] Primary culture fluid: Add glutamine, fetal bovine serum and antibiotics to the M199 culture fluid, so that the concentration of glutamine is 600 μg / ml, fetal bovine serum accounts for 25% of the total volume, and the concentration of penicillin is 100 IU / ml. The streptomycin concentration was 100 μg / ml, the streptomycin sulfate concentration was 40 μg / ml, and the amphotericin B concentration was 40 μg / ml.

[0030] Subculture medium: Add glutamine, fetal b...

Embodiment 2

[0046] 1. Prepare PBS disinfectant and cell culture medium

[0047] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 300IU / ml, the concentration of streptomycin is 300μg / ml, the concentration of streptomycin sulfate is 20μg / ml, and the concentration of amphotericin B is 20μg / ml.

[0048] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 400 μg / ml, and the fetal bovine serum accounts for 10% of the total volume.

[0049] Primary culture solution: Add glutamine, fetal bovine serum and antibiotics to the M199 culture medium, so that the concentration of glutamine is 800 μg / ml, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200 IU / ml. The streptomycin concentration was 30 μg / ml, the streptomycin sulfate concentration was 20 μg / ml, and the amphotericin B concentration was 20 μg / ml.

[0050] Subculture medium: Add glutamine, feta...

Embodiment 3

[0061] 1. Prepare PBS disinfectant and cell culture medium

[0062] PBS disinfectant: Add antibiotics to PBS so that the concentration of penicillin is 300IU / ml, the concentration of streptomycin is 300μg / ml, the concentration of streptomycin sulfate is 20μg / ml, and the concentration of amphotericin B is 20μg / ml.

[0063] Basal culture medium: add glutamine and fetal bovine serum to the M199 medium, so that the concentration of glutamine is 400 μg / ml, and the fetal bovine serum accounts for 10% of the total volume.

[0064] Primary culture solution: Add glutamine, fetal bovine serum and antibiotics to the M199 culture medium, so that the concentration of glutamine is 800 μg / ml, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200 IU / ml. The streptomycin concentration was 30 μg / ml, the streptomycin sulfate concentration was 20 μg / ml, and the amphotericin B concentration was 20 μg / ml.

[0065] Subculture medium: Add glutamine, feta...

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Abstract

The invention relates to a method for constructing a sinocyclocheilus rhinocerous heart cell line. The method comprises the following steps: 1, obtaining of sinocyclocheilus rhinocerous heart tissues, wherein sinocyclocheilus rhinocerous is sterilized through combination of penicillin and ethyl alcohol; 2, primary culture, wherein a culture solution M199 which contains glutamine, penicillin, streptomycin, streptomycin sulphate and amphotericin is adopted for culture, and the osmotic pressure of the culture solution is 280-330 mOsm / l; 3, secondary culture, wherein when the tenth generation is reached, cell culture fluid is replaced with base culture fluid, and the sinocyclocheilus rhinocerous heart cell line is constructed successfully. According to the method for constructing the sinocyclocheilus rhinocerous heart cell line, the obtained sinocyclocheilus rhinocerous heart cell line is in the fibrous form and can conduct continuous passage and be directly applied to research of biological characteristics, requirements of genetic resource preservation and theoretical research conducted on the rare species, namely the sinocyclocheilus rhinocerous, can be satisfied, meanwhile, the heart cell line is the first heart cell line of cave fishes in China, and the foundation is laid for cave adaptability research at the cellular level.

Description

technical field [0001] The invention relates to a method for establishing a cell line by using the heart tissue of the rhino horned barb, and belongs to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. Background technique [0002] Sinocyclocheilus rhinocerous (Regan, 1904) belongs to the genus Sinocyclocheilus of Cypriniformes (Cypriniformes) Cyprinidae. It is a new cave species published in 1994. At present, the known distribution of this species is The point is the underground river in Huancheng Township, Luoping County, Yunnan Province, which belongs to the Nanpanjiang River system. The research on it is only found in the description of its taxonomic features, the confirmation of its taxonomic status and its phylogenetic relationship. [0003] Due to the small population of rhino-horned barb and the difficulty of obtaining it, how to protect and preserve the germplasm resources of this rare cave fish has become a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A01N1/02C12R1/91
Inventor 王晓爱潘晓赋杨君兴杨坤凤刘倩
Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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