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Chicken Marek's disease vaccine produced by using continuous passage cell line and production method

A technology for Marek's disease and subculture cells, which is applied in the field of chicken Marek's disease vaccine production by quail sarcoma fibroblast QT6, to achieve the effect of small batch-to-batch difference, increased virus yield, and stable immune effect

Inactive Publication Date: 2013-06-19
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the development of many avian or mammalian cell lines (e.g. EP0748867 and EP0884382), there is still no continuous cell line as an effective substitute for chicken embryo fibroblasts (CEF) in MD vaccines

Method used

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  • Chicken Marek's disease vaccine produced by using continuous passage cell line and production method
  • Chicken Marek's disease vaccine produced by using continuous passage cell line and production method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The method of embodiment 1 passage cell production chicken Marek's disease vaccine

[0036] (1) Subculture and cultivation of QT6 cells for seedling preparation:

[0037] QT6 cells were digested and passaged with EDTA-trypsin cell dispersion solution, one bottle was passed to three bottles, and the cell growth medium was used to continue culture at 37°C, and one generation was carried out every 4 days. When a good monolayer is formed, it can be used for further passage or virus inoculation. The cell seeding density was 2×10 5 cells / ml;

[0038] (2) Propagation of cytotoxic species:

[0039] Take the chicken Marek's virus CVI988 / Rispens strain preserved in liquid nitrogen, rejuvenate and subculture on the above-mentioned cells for 1 to 2 generations, and prepare working seed virus;

[0040] (3) Vaccination of working viruses:

[0041] Take a 10,000ml cell spinner bottle that has formed a good monolayer, discard the cell growth medium, inoculate 1,000ml of maintenanc...

Embodiment 2

[0059] Example 2 Comparison of Chicken Marek's Disease Vaccine and the Use of CEF Cells to Prepare Chicken Marek's Disease Vaccine

[0060] 1. The main steps of preparing existing chicken Marek's disease vaccine are as follows:

[0061] (1) Preparation of chicken embryo fibroblast (CEF) monolayer:

[0062] Select 10-day-old well-developed SPF chicken embryos, first use iodine tincture cotton balls to sterilize the air chamber, and then use 75% alcohol cotton balls to deiodine, aseptically pick out the embryo bodies and place them in sterile plates, and use 0.1M PBS Wash the embryo body and remove the eyes, claws and viscera, then prepare the CEF suspension by continuous digestion method to make a cell density of 1.5 million / ml, mix well and inoculate the cell culture flask. ;

[0063] (2) Propagation of cytotoxic species:

[0064] Take the chicken Marek's virus CVI988 / Rispens strain in Example 1, rejuvenate and subculture on the above-mentioned CEF for 1 to 2 generations, a...

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Abstract

The invention discloses a production method for a chicken Marek's disease vaccine by using a continuous passage cell line. The method comprises the following steps: (1) passage and culture of a cell used for vaccine preparation; (2) propagation of a cellular virus seed; (3) inoculation of a virus; (4) centrifugal extraction of the cell for collection of the virus; and (5) distribution and split charging of the vaccine. According to the invention, passage cells are used to replace original primary chicken embryonic fibroblasts for production of the chicken Marek's disease vaccine, so production cost is substantially reduced; the production method has a simple process, is stable and is easy to operate; virus plaque content is high, the vaccines of different batches have small difference, vaccine quality is easily controllable, and output and quality of the vaccine can be substantially improved. The chicken Marek's disease vaccine produced in the invention has high immune efficacy on a standard virulent strain and has an immune effect superior to that of a product of a same kind.

Description

technical field [0001] The invention relates to a chicken Marek's disease vaccine produced by continuous passage cell line and a method thereof, in particular to a chicken Marek's disease vaccine produced by using quail sarcoma fibroblast QT6 and a method thereof. Background technique [0002] Marek's disease (Marek's disease, MD) is a contagious neoplastic disease of chickens caused by Marek's disease virus (MDV) of the herpesviridae family, characterized by lymphoid tissue hyperplasia and tumor formation, in the peripheral Infiltration of mononuclear cells occurs in nerves, gonads, various organs of iris, muscles and skin. While MD causes tumors and death clinically, more importantly, it damages the immune organs of infected chickens, resulting in immunosuppression, resulting in decreased body resistance and reduced or no immune response to vaccination, causing complications of other diseases And secondary infection, causing huge economic losses. [0003] Vaccination is ...

Claims

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Application Information

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IPC IPC(8): A61K39/255C12N7/00A61P31/22
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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