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A grass carp swim bladder epithelioid cell line and its 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: 2016-07-06
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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  • A grass carp swim bladder epithelioid cell line and its application
  • A grass carp swim bladder epithelioid cell line and its application
  • A grass carp swim bladder epithelioid cell line and its 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 and live 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 200ulPBS), and cut it with surgical scissors. Cut into pieces; use a...

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] To compare the proliferation of two strains of GCRV virus (GCRVJX-0901, GCRVHZ08) in different cells: according to the above method, a specific concentration of virus (GCRVJX-0901160 copies / μl, GCRVHZ083.6×10 6copies / μl) were inoculated in the 50th generation grass carp swim bladder cells (GSB-E, GSB-F) of the present invention and other five commonly used fish cell lines: CIK (grass carp kidney cells, purchased from China Center for Type Culture Coll...

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Abstract

The invention discloses a grass carp swim bladder epithelioid cell line and its application. The grass carp swim bladder epithelial-like cell line with the preservation number CCTCC No.C201443, the proliferation of grass carp reovirus GCRV in this cell line is significantly higher than that of CIK, CO, PSF, FHM, EPC and other cell lines commonly used to isolate GCRV at present , and the cell lesion time is fast, therefore, this cell line can be well applied to the isolation and propagation of grass carp reovirus and the research of grass carp haemorrhagic 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( Ctenopharyngodonidellus ) 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 reovirus (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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Patent Type & Authority Patents(China)
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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