Method for preparing haemorrhagic fever vaccine by culturing primary cells in multilayer culture flask

A multi-layer culture and original culture technology, which is applied in the direction of virus antigen components, recovery/purification, inactivation/attenuation, etc., can solve the problems of small cell culture area, large greenhouse space occupation, high labor intensity, etc. The effect of fewer cells, less space, and more cells

Active Publication Date: 2012-03-14
ZHEJIANG PUKANG BIOTECH
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Problems solved by technology

The disadvantages of spinner bottle culture are: small cell culture area, large greenhouse space, high labor intensity and easy contamination

Method used

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  • Method for preparing haemorrhagic fever vaccine by culturing primary cells in multilayer culture flask
  • Method for preparing haemorrhagic fever vaccine by culturing primary cells in multilayer culture flask
  • Method for preparing haemorrhagic fever vaccine by culturing primary cells in multilayer culture flask

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Embodiment Construction

[0014] Below in conjunction with accompanying drawing and embodiment the invention will be further described:

[0015] The method for preparing hemorrhagic fever vaccine by cultivating primary cells in multilayer culture flasks is as follows:

[0016] Select 10-20 day-old clean-grade gerbils, fumigate them to death with ether, wash and disinfect the mouse corpses; take out the kidneys aseptically, cut them into pieces, digest them with trypsin, and disperse them in MEM culture solution containing 3-5% calf serum Cells, prepare cell suspension, pack into multi-layer cell culture flasks, 8000ml per bottle, culture at 37±1°C for 2-3 days, then replace with MEM culture medium containing 1% calf serum.

[0017] After 4-5 days, the cells were covered with a dense monolayer, and the virus was inoculated at a MOI of 0.2 (every 100cm 2 Inoculate 1ml of hemorrhagic fever virus working seeds with a titer of 6.0lgCCID50 / ml), culture at 34.5±1°C for 22-26 hours, discard the culture medium...

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Abstract

The invention relates to a method for preparing a haemorrhagic fever vaccine by culturing primary cells in a multilayer culture flask. The method comprises the following steps: anatomizing an animal to obtain the primary cells; inoculating half or one third cell of cell amount required by a spinner flask into the multilayer cell culture flask, adding proper culture solution in a hot house for culturing; and inoculating virus for culturing after the cell grows full of a single layer, obtaining virus culture solution, and preparing the vaccine after purification. The multilayer cell flask has the advantages of small amount of inoculated primary cells, large amount of the cultured cells, small occupied space, no need of a flask spinning machine and high yield.

Description

Technical field [0001] The invention belongs to the cultivation of primary cells and virus propagation, and mainly relates to a method for preparing hemorrhagic fever vaccines by culturing primary cells in multilayer culture flasks. Background technique [0002] Cell culture (Cell culture) is to simulate the physiological conditions in the body, take the cells out of the body, make them survive, grow, reproduce and pass on under artificial conditions, and carry out the research and production of cell life process, cell carcinogenesis, cell engineering and other issues vaccine. The tissue or cells directly removed from the human or animal body for culture are called primary culture. The time of primary cultured cells in vitro is short, and their properties are similar to those in vivo. Generally speaking, tissues and cells in immature state, such as: animal embryos, organs of young, etc., are easier to carry out primary culture. Traditional vaccine preparation mostly adopt...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12A61P31/14C12N7/06C12N7/02
Inventor 姚伟钟建琴苏波
Owner ZHEJIANG PUKANG BIOTECH
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