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A kind of construction method of barb barb fin cell line

A technology of barb barb and construction method, which is applied in cell dissociation methods, artificial cell constructs, animal cells, etc., can solve problems such as no similar reports, and achieve simple and easy operation, strong repeatability, and high cell volume. big effect

Active Publication Date: 2020-03-31
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 kind of construction method of barb barb fin cell line
  • A kind of construction method of barb barb fin cell line
  • A kind of construction method of barb barb fin cell line

Examples

Experimental program
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Effect test

Embodiment 1

[0031] 1. Preparation of HBSS disinfection solution and cell culture solution

[0032] HBSS disinfectant solution: Add antibiotics to HBSS so that the concentration of penicillin is 300 IU / mL, the concentration of streptomycin is 300 μg / mL, the concentration of gentamicin is 40 μg / mL, and the concentration of amphotericin B is 40 μg / mL.

[0033] Basal culture medium: add cell growth factors and fetal bovine serum to the DMEM / F12 medium, so that the concentration of cell growth factors is 8 ng / L, and the fetal bovine serum accounts for 15% of the total volume.

[0034] Primary culture medium: add cell growth factors, fetal bovine serum and antibiotics to DMEM / F12 medium, so that the concentration of cell growth factors is 12ng / L, fetal bovine serum accounts for 20% of the total volume, and the concentration of penicillin is 300IU / L mL, the concentration of streptomycin was 300 μg / mL, the concentration of gentamicin was 50 μg / mL, and the concentration of amphotericin B was 50 μg...

Embodiment 2

[0046] 1. Preparation of HBSS disinfection solution and cell culture solution

[0047] HBSS disinfectant: Add antibiotics to HBSS so that the concentration of penicillin is 400 IU / mL, the concentration of streptomycin is 400 μg / mL, the concentration of gentamicin is 30 μg / mL, and the concentration of amphotericin B is 30 μg / mL.

[0048] Basal culture medium: add cell growth factors and fetal bovine serum to the L-15 medium, so that the concentration of cell growth factors is 10 ng / L, and the fetal bovine serum accounts for 15% of the total volume.

[0049] Primary culture medium: add cell growth factors, fetal bovine serum and antibiotics to the L-15 medium, so that the concentration of cell growth factors is 10ng / L, fetal bovine serum accounts for 20% of the total volume, and the concentration of penicillin is 300IU / L mL, the concentration of streptomycin was 300 μg / mL, the concentration of gentamicin was 40 μg / mL, and the concentration of amphotericin B was 40 μg / mL.

[005...

Embodiment 3

[0061] 1. Preparation of HBSS disinfection solution and cell culture solution

[0062] HBSS disinfectant solution: Add antibiotics to HBSS so that the concentration of penicillin is 200 IU / mL, the concentration of streptomycin is 200 μg / mL, the concentration of gentamicin is 60 μg / mL, and the concentration of amphotericin B is 60 μg / mL.

[0063] Basal culture medium: add cell growth factors and fetal bovine serum to the DMEM / F12 medium, so that the concentration of cell growth factors is 10 ng / L, and the fetal bovine serum accounts for 10% of the total volume.

[0064] Primary culture medium: add cell growth factors, fetal bovine serum and antibiotics to the DMEM / F12 medium, so that the concentration of cell growth factors is 12ng / L, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200IU / L mL, the concentration of streptomycin was 200 μg / mL, the concentration of gentamicin was 30 μg / mL, and the concentration of amphotericin B was ...

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Abstract

The invention relates to a construction method of a cell line of a fin of spinibarbus hollandi. The construction method comprises the following steps of 1), acquisition of the fin of the spinibarbus hollandi: jointly sterilizing a fish body by adopting methylene blue and ethyl alcohol; 2), primary culture: carrying out the culture by adopting a DMEM (Dulbecco's Modified Eagle Media) / F12 or L-15 culture solution containing a cell growth factor, fetal bovine serum, penicillin, streptomycin, gentamicin and amphotericin B; 3), subculture: when handing to an eighth generation, changing a cell culture solution to a basal culture solution, so that the cell line of the fin of the spinibarbus hollandi is established successfully. The cell line of the fin of the spinibarbus hollandi, which is obtained through the construction method provided by the invention, can be used for realizing continuous passage, is directly applied to biological characteristic research, can be used for meeting the demands of the germplasm-resource storage and the theoretical research of an economic species of the spinibarbus hollandi, can also be repeatedly freeze-thawed, and can be applied to the important research of cell biology, virology, toxicology, molecular biology, genetics, immunology and the like.

Description

technical field [0001] The invention relates to a method for establishing a cell line by using light barb fin tissue, and belongs to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. [0002] technical background [0003] Spinibarbus hollandi, commonly known as green bamboo carp, green stick, kengjian, light eye fish, yellow juan, rough scale fish, belongs to Cyprinidae (Cyprinidae) subfamily Barbinae (Spinibarbus) . Historically, the barbed barb was the main target of fishermen in several major water systems in China, with the largest individual weighing more than 25 kilograms. It is mainly distributed in the Yangtze River, Qiantang River, Minjiang River, Jiulong River, Pearl River, Yuanjiang River, Taiwan Island and Hainan Island. According to the survey of the Red River water system in China in recent years, the population decline of the barbed barb is very serious, and its population has been occasionally seen in...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A01N1/02
CPCA01N1/021C12N5/0602C12N2500/84C12N2501/10C12N2509/00
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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