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Anguilla anguilla liver cell line and construction method and application thereof

A technology of liver cells and construction method, applied in the field of marine fish cell culture, can solve problems such as failure to meet research needs, unreasonable aquaculture layout, and decreased immunity of fish, achieve simple and clear screening conditions, and simple and easy technical solutions , good effect of sticking to the wall

Inactive Publication Date: 2015-04-22
BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the rapid development of the aquaculture industry, problems such as unreasonable aquaculture layout, excessive density, and poor water flow have commonly occurred, coupled with the situation of feeding deteriorating bait and abusing drugs from time to time, resulting in the decline of fish immunity and water pollution, various diseases Frequent outbreaks, many of which are viral cases, and the existing European eel cell line models are very few, which are far from meeting the needs of research

Method used

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  • Anguilla anguilla liver cell line and construction method and application thereof
  • Anguilla anguilla liver cell line and construction method and application thereof
  • Anguilla anguilla liver cell line and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0035] Example 1: Primary culture of European eel liver cells

[0036] Experimental animals:

[0037] Healthy juvenile European eel with a body length of about 20cm and a weight of about 20g.

[0038] Reagent:

[0039] L-15 (purchased from Hyclone); 1000000U / 10mg penicillin-streptomycin mixed solution, absolute ethanol, tincture of iodine (purchased from Shanghai Sangong); NaCl, Na2HPO 4 , KCl, KH2PO 4 , NaHCO 3 , trypsin (Trypsin) (purchased from Sigma); fetal bovine serum (Fetal bovine serum, FBS) (purchased from Gibco).

[0040] instrument:

[0041] Ultra-clean bench (Air Tech); inverted fluorescence microscope (Nikon); biochemical incubator (Boxun); ultra-pure water machine (Millipore).

[0042] Consumables:

[0043] Ophthalmic scissors, ophthalmic tweezers, and Venus scissors for dissection (Liuliu Ophthalmology); 25mL, 50mL cell culture flasks (BD Falcon); 15mL centrifuge tubes (BD Falcon); 5mL elbow glass dropper.

[0044] step:

[0045] 1) Pre-test p...

example 2

[0052] Example 2: Early subculture of European eel liver cells

[0053] Reagent:

[0054] L-15 (purchased from Hyclone); 1000000U / 10mg penicillin-streptomycin mixture (purchased from Shanghai Sangong); NaCl, Na2HPO 4 , KCl, KH2PO 4 , NaHCO 3 , trypsin (Trypsin) (purchased from Sigma); fetal bovine serum (Fetal bovine serum, FBS) (purchased from Gibco)

[0055] instrument:

[0056] Ultra-clean bench (Air Tech); inverted fluorescence microscope (Nikon); biochemical incubator (Boxun); ultra-pure water machine (Millipore); centrifuge (Beckman)

[0057] Consumables:

[0058] 25mL, 50mL cell culture flask (BD Falcon); 15mL centrifuge tube (BD Falcon); 5mL elbow glass dropper

[0059] step:

[0060] 1) Preparation of complete culture medium for primary and early subculture: prepare L-15 medium as the basis, containing fetal bovine serum at a final concentration of 15%, penicillin at a final concentration of 200U / mL, and a final concentration of 200μg / mL mL of strept...

example 3

[0064] Example 3: The routine subculture of European eel liver cells and the establishment of cell lines:

[0065] Reagent:

[0066] L-15 (purchased from Hyclone); glycerin, methanol, glacial acetic acid, Giemsa powder (purchased from Shanghai Sangong); NaCl, Na2HPO 4 , KCl, KH2PO 4 , NaHCO 3 , trypsin (Trypsin) (purchased from Sigma); fetal bovine serum (Fetal bovine serum, FBS) (purchased from Gibco)

[0067] instrument:

[0068] Ultra-clean bench (Air Tech); inverted fluorescence microscope (Nikon); biochemical incubator (Boxun); ultra-pure water machine (Millipore)

[0069] Liquid nitrogen tank (Locator), centrifuge (Beckman)

[0070] Consumables:

[0071] 50mL cell culture flask (BD Falcon), 250mL cell culture flask (Corning); 15mL centrifuge tube (BD Falcon); glass slide; 5mL elbow glass dropper; standard glass destaining dish

[0072] step:

[0073] 1) Preparation of complete culture medium for routine subculture: prepare a complete culture medium for...

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Abstract

The invention discloses an anguilla anguilla liver cell line and a construction method and application thereof, and belongs to the technical field of sea fish cell culture. The cell line is preserved in the Chinese typical culture preservation center on December 16, 2014, and the serial number of preservation registration is CCTCC NO: C2014239. The anguilla anguilla liver cell line is fibroblast cells, the continuous passage achieves more than 60 generations, the adherence after cryopreservation resuscitation is good, the growth is stable, the liver cell line can be directly used for research on aquatic virology, anguilla anguilla cell biological characteristics and immune resistance gene function, and the lever cell line has been proved to be sensitive to eels herpes virus. According to the anguilla anguilla liver cell line and the construction method and application thereof, the constructing method is simple and convenient to implement, the repeatability is high, and the cost is moderate. The anguilla anguilla liver cell line and the construction method and application thereof can also be applied to the preparation of other anguilla anguilla cell lines.

Description

technical field [0001] The present invention relates to a kind of European eel ( Anguilla Anguilla ) liver cell line and its construction method and application, belonging to the technical field of seawater fish cell culture. Background technique [0002] Fish cell culture began in the 1960s. Since Wolf et al. established the first permanent teleost cell line—the rainbow trout gonad cell line RTG-2 in 1962, more than 250 fish cell lines have been established at home and abroad. More than 40 kinds of viruses have been successfully isolated by cell culture, and the success of fish embryonic stem cell culture has effectively promoted the development of genetic engineering. Fish research experiments have high requirements for the maintenance of breeding sites, equipment and water environment. Using fish culture cells as an experimental model has advantages that live fish cannot match: (1) Low cost, and the maintenance of cell lines does not require large (2) The repeatability ...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00C12R1/91C12R1/93
Inventor 郑在予杨金先龚晖葛均青林天龙
Owner BIOLOGICAL TECH INST OF FUJIAN ACADEMY OF AGRI SCI
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