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Grass carp swim bladder epithelioid cells line and application

A cell line and grass carp technology, applied in the field of cytology, can solve the problems of grass carp swim bladder cell lines that have not been reported

Inactive Publication Date: 2014-07-30
广州普麟生物制品有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] So far, there are grass carp kidney cell lines, grass carp rostral tissue cell line ZC-7901 and its substrain ZC-7901S1, rostral fibroblast cell line PSF, grass carp caudal fin tissue diploid cell line, grass carp blastula cell line, Several cell lines such as the liver cell line have been reported in the literature, but there is no report on the grass carp swim bladder cell line

Method used

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  • Grass carp swim bladder epithelioid cells line and application
  • Grass carp swim bladder epithelioid cells line and application
  • Grass carp swim bladder epithelioid cells line and application

Examples

Experimental program
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Effect test

Embodiment 1

[0018] 1. Preparation of cell culture medium

[0019] Take the M199 medium of GIBCO company, add fetal bovine serum accounting for 10-20% of the total volume of the medium (the percentages mentioned below are all volume percentages unless otherwise specified), and the final concentration is 20ng / ml basic fibroblast growth factor (bFGF), epidermal growth factor (EGF) with a final concentration of 1ng / ml, stored at 4°C, and used for later use.

[0020] 2. Primary culture

[0021] The 4 grass carps (10±0.5cm) used in the present invention are from the breeding base of the Pearl River Fisheries Research Institute, about 40 days old, and the primary cells were separated and cultured in July 2011.

[0022] Soak fresh grass carp in 75% alcohol for 1-2 minutes, take out the swim bladder aseptically, remove the membrane with tweezers, rinse twice with PBS, put it in a 5ml sterile penicillin vial (containing 200ul PBS), and cut it with surgical scissors. Shred it; use a pipette to mov...

Embodiment 2

[0034] Detect the sensitivity of grass carp hemorrhagic disease virus (GCRV) to different cell lines: wait until the cells are inoculated to 25cm 2 After the bottom wall of the cell flask, put 1mL virus solution (3.2×10 3 copy / μl) into the cell culture flask, after 1 hour, remove the virus solution, replace with fresh cell culture medium containing 3% fetal bovine serum, and continue the culture. After about 2 days, after the cytopathic effect (CPE) is observed, the The cells were observed under the electron microscope.

[0035] Compare the proliferation of two strains of GCRV virus (GCRV JX-0901, GCRV HZ08) in different cells: mix specific concentrations of virus (GCRV JX-0901 160 copies / μl, GCRV HZ08 3.6×10 6copies / μl) were inoculated in the 50th generation grass carp swim bladder cells (GSB-E, GSB-F) of the present invention and other 5 commonly used fish cell lines: CIK (grass carp kidney cells, purchased from China Center for Type Culture Collection) , PSF (grass carp r...

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PUM

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Abstract

The invention discloses a grass carp swim bladder epithelioid cells line and an application. According to the grass carp swim bladder epithelioid cells line with a preservation number of CCTCC No.C201443, Grass Carp Reovirus GCRV propagation amount in the cell line is obviously higher than that of the cell lines such as CIK, CO, PSF, FHM and EPC used for separating GCRV, cell pathology time is fast, and the cell line can better used in the researches of separation and breeding of Grass Carp Reovirus and grass carp hemorrhagic disease vaccine.

Description

technical field [0001] The invention relates to the technical field of cytology, in particular to a grass carp swim bladder epithelial cell line and application thereof. Background technique [0002] grass carp( Ctenopharyngodon idellus ) belongs to the genus Grass carp of Cyprinidae Cyprinidae, a herbivorous fish with low feeding cost and wide breeding area. Breeding is also one of the main freshwater species in Guangdong Province. However, disease problems are prone to occur during grass carp culture, among which viral hemorrhagic disease is the worst, causing huge losses to my country's grass carp culture industry. Grass carp hemorrhagic disease caused by Grass carp reovims (GCRV) infection has become one of the important factors restricting the healthy development of grass carp industry. At present, there is no specific and effective drug and method for the prevention and treatment of grass carp haemorrhagic disease, and the most effective method is still immune preve...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00C12R1/91
Inventor 王英英王庆曾伟伟吴淑勤梁红茹李莹莹石存斌
Owner 广州普麟生物制品有限公司
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