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Pig tonsil cell line susceptible to JEV

A cell line and tonsillar technology, applied in the direction of genetically modified cells, cells modified by introducing foreign genetic material, animal cells, etc., to achieve the effect of expanding the scope

Inactive Publication Date: 2019-11-19
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a kind of porcine tonsil cell line susceptible to JEV, this method solves the problem that there is no porcine tonsil cell line infected with porcine virus at present, it has the function of infinite passage, and can maintain the porcine tonsil cell Features, can maintain normal karyotype during the passage process, there is no change in chromosome structure and number, and is sensitive to JEV, can be used for the passage of JEV and related research

Method used

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  • Pig tonsil cell line susceptible to JEV
  • Pig tonsil cell line susceptible to JEV
  • Pig tonsil cell line susceptible to JEV

Examples

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experiment example

[0028] Characterization of JEV-susceptible porcine tonsil cell line by experimental example

[0029] Using immunofluorescence to verify the characteristics of JEV-susceptible porcine tonsil cell lines, the specific steps are as follows:

[0030] (1) Cell slide

[0031] Take the glass slides in a 24-well plate, add 1 mL of DMEM / F12 medium to each well, and add 0.02 million cells / well. Place in the incubator for 2 hours or overnight.

[0032] (2) Cells were inoculated with 5MOI of JEV virus, DMEM medium with 2% FBS was replaced two hours later, and cell samples were harvested at different time points.

[0033] (3) Fixed

[0034] At different time points after the cells were sliced, the medium was aspirated, washed once with PBS (phosphate buffered saline), and fixed with 4% PFA (paraformaldehyde) at 4°C for 30 min. Wash with PBS 3×5min / time.

[0035] (4) Membrane closure

[0036] Remove moisture from the glass slide, place it on the petri dish support, glass slide sealing ...

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Abstract

The invention discloses a pig tonsil cell line susceptible to JEV. The cell line infects a primary pig tonsil cell line through lentivirus particles, cell screening is conducted in a serum-containingDMEM / F12 culture solution in which puromycin is added, positive cells are subjected to enlarged culture through a culture solution not containing puromycin, and the pig tonsil cell line is obtained. The lentivirus particles are obtained by taking pHY-sv4O-Puro as a lentivirus carrier and taking psPAX2 and pCMV-VSV-G as an auxiliary packaging carrier for co-transfection to enter 293T cells. The cell line has the infinite passage function, is susceptible to JEV and can be used for related research.

Description

technical field [0001] The invention relates to a porcine tonsil cell line, in particular to a porcine tonsil cell line susceptible to JEV. Background technique [0002] Pig Japanese encephalitis (referred to as JE for short) is a common viral encephalitis caused by porcine epidemic encephalitis virus (Japanese encephalitis virus, JEV). Both poultry and humans can be infected by the disease. The disease is mainly transmitted through the bite of blood-sucking insects such as mosquitoes, and pigs of any breed and sex are susceptible to the disease. The clinical symptoms are mainly abortion or stillbirth in pregnant sows, encephalitis, ataxia, fever, and death in newborn piglets, obvious swelling of testicles in boars, neurological symptoms in individual sick pigs, and some pregnant animals may also be infected and aborted . [0003] PK-15 cells (porcine kidney epithelial cells), ST cells (porcine testicular cells), PIEC cells (porcine hip arterial endothelial cells) are all...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12R1/91
CPCC12N5/0634C12N2501/998C12N2510/00
Inventor 马志永刘珂郗淑敏邵东华魏建超李蓓蓓邱亚峰
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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