Sinocyclocheilus anshuiensis spleen cell line construction method

A technology of Anshui golden thread barb and construction method, which is applied in artificial cell constructs, microorganism-based methods, animal cells, etc., can solve problems such as no similar reports, and achieve simple and easy operation, large cell volume, and stability. good effect

Active Publication Date: 2016-02-17
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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  • Sinocyclocheilus anshuiensis spleen cell line construction method
  • Sinocyclocheilus anshuiensis spleen cell line construction method
  • Sinocyclocheilus anshuiensis spleen cell line construction method

Examples

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Embodiment 1

[0024] 1. Preparation of HBSS disinfection solution and cell culture solution

[0025] HBSS disinfectant solution: Add antibiotics to HBSS so that the concentration of penicillin is 200IU / ml, the concentration of streptomycin is 200μg / ml, the concentration of gentamicin is 30μg / ml, and the concentration of amphotericin B is 30μg / ml.

[0026] Basal culture medium: Add glutamine and fetal bovine serum to the L-15 medium, so that the concentration of glutamine is 360 mg / L, and the fetal bovine serum accounts for 15% of the total volume.

[0027] Primary culture medium: Add glutamine, fetal bovine serum and antibiotics to the L-15 medium so that the concentration of glutamine is 700mg / L, fetal bovine serum accounts for 20% of the total volume, and the concentration of penicillin is 150IU / L ml, the concentration of streptomycin is 150 μg / ml, the concentration of gentamicin is 30 μg / ml, and the concentration of amphotericin B is 30 μg / ml.

[0028] Subculture medium: add glutamine, ...

Embodiment 2

[0039] 1. Preparation of HBSS disinfection solution and cell culture solution

[0040] HBSS disinfectant: Add antibiotics to HBSS so that the concentration of penicillin is 100IU / ml, the concentration of streptomycin is 100μg / ml, the concentration of gentamicin is 40μg / ml, and the concentration of amphotericin B is 40μg / ml.

[0041] Basal culture medium: Add glutamine and fetal bovine serum to the L-15 medium, so that the concentration of glutamine is 400 mg / L, and the fetal bovine serum accounts for 15% of the total volume.

[0042] Primary culture medium: Add glutamine, fetal bovine serum and antibiotics to the L-15 medium, so that the concentration of glutamine is 600mg / L, fetal bovine serum accounts for 20% of the total volume, and the concentration of penicillin is 100IU / L ml, the concentration of streptomycin is 100 μg / ml, the concentration of gentamicin is 40 μg / ml, and the concentration of amphotericin B is 40 μg / ml.

[0043] Subculture medium: add glutamine, fetal bo...

Embodiment 3

[0055] 1. Preparation of HBSS disinfection solution and cell culture solution

[0056] 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 20 μg / ml, and the concentration of amphotericin B is 20 μg / ml.

[0057] Basal culture medium: Add glutamine and fetal bovine serum to the L-15 medium to make the concentration of glutamine 300 mg / L, and the fetal bovine serum accounts for 10% of the total volume.

[0058] Primary culture solution: Add glutamine, fetal bovine serum and antibiotics to the L-15 culture medium, so that the concentration of glutamine is 600 mg / L, fetal bovine serum accounts for 15% of the total volume, and the concentration of penicillin is 200 IU / L ml, the concentration of streptomycin is 200 μg / ml, the concentration of gentamicin is 30 μg / ml, and the concentration of amphotericin B is 30 μg / ml.

[0059] Subculture medium: ad...

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Abstract

The invention relates to a Sinocyclocheilus anshuiensis spleen cell line construction method. The construction method comprises the following steps: (1) obtaining a Sinocyclocheilus anshuiensis spleen: directly disinfecting a fish body by adopting ethyl alcohol; (2) performing primary culture: culturing by adopting B-15 culture solution containing glutamine, fetal calf serum, penicillin, streptomycin, gentamicin and amphotericin B; (3) subculturing: when culturing to the eighth generation, changing the cell culture solution into basic culture solution, thus successfully constructing a Sinocyclocheilus anshuiensis spleen cell line. The Sinocyclocheilus anshuiensis spleen cell line obtained by the construction method can realize continuous passage, is directly applied to biological characteristic research, and can meet the requirements of germplasm resource conservation and theoretical research on Sinocyclocheilus anshuiensis as a rare species; more importantly, the cell line is the first cell line of cave fish in China, and lays the foundation for the cellular level cave adaptability research.

Description

technical field [0001] The invention relates to a method for establishing a cell line by using the spleen tissue of Anshui goldenrod barb, and belongs to the technical field of freshwater aquatic organism cell culture and ultra-low temperature cryopreservation. Background technique [0002] Anshui golden barb Sinocyclocheilus anshuiensis (Regan, 1904) belongs to Cypriniformes (Cypriniformes) Cyprinidae (Cyprinidae) golden line barb genus (Sinocyclocheilus), is a new species of blind fish published in 2013, currently, the known species of this species The distribution point is only a cave in Anshui Village, Luolou Town, Lingyun County, Guangxi Zhuang Autonomous Region, and the research on it can only be found in the description of its taxonomic characteristics and the confirmation of its taxonomic status. [0003] Because the population of Anshui goldenrod is small and difficult to obtain, 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 Applications(China)
IPC IPC(8): C12N5/071A01N1/02C12R1/91
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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