Method for culturing primary cells
A primary cell and culture method technology, applied in the field of primary cell culture, can solve problems such as difficult quality control, high shear force, and cell culture technology that cannot completely eliminate exogenous factor contamination, so as to improve product quality and production scale and production efficiency, quality improvement, and the effect of increasing cell culture rate
Inactive Publication Date: 2010-12-29
LIAONING ANDI BIO TECH
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Problems solved by technology
[0003] However, this technology is mainly used for the cultivation of passage cell lines and for primary cell culture. Due to the high shear force, its large-scale production application is limited.
[0004] Primary cells can be used to produce rabies vaccine, Japanese encephalitis vaccine, measles virus vaccine, mumps virus vaccine and epidemic hemorrhagic fever virus vaccine, etc. The production of these vaccines requires the use of a large number of animals, and traditional cell culture techniques cannot completely exclude foreign sources The possibility of factor contamination makes it difficult to control the quality and restricts the large-scale production of vaccine products
Method used
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Experimental program
Comparison scheme
Effect test
Embodiment 1
[0014] Example 1: Hamster kidney cells were cultured using a bioreactor and using polyester slices as carriers
[0015] 1. The kidney cells were dissected from the hamster and digested with trypsin to obtain primary cells;
[0016] 2. Take 1×10 4 ~1×10 7 The density of cells / ml was inoculated in the bioreactor equipped with polyester chip carrier;
[0017] 3. After inoculation, keep the stirring speed between 50-120rpm, and cultivate at 30°C-38°C for 5-25 days. During the culture period, add fresh medium to keep the glucose content in the medium at 2-2.5g / Lift. Adjust the flow rate of O2, N2, CO2 gas, and detect during the incubation period. Lactose concentration was measured by lactose dehydrogenase method; glucose concentration was measured by glucose oxidase method, and the growth status of primary animal cells was determined by glucose consumption.
[0018] The features of the present invention are:
[0019] The cells used for culture can be primary animal cells suc...
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The invention provides a method for culturing primary cells. The problem to be solved is that: the primary cells can be used for producing a large number of vaccines such as a rabies vaccine and the like; and the production of the vaccines needs to use a large number of animals, while conventional cell culture technology cannot completely rule out the possibility of exogenous factor pollution, so the quality is difficult to control and the large-scale production of vaccine products is restricted. The main point of the method is that: obtained primary cells are cultured by taking a polyester chip carried by a bioreactor as a carrier. The method has the positive effects that: each gram of the polyester chip carrier is equivalent to a 3L-transfer bottle of which the inner surface is about 1,160 square centimeters; and the cell density is up to 10<8> cells / milliliter. Analyzed and compared by synthesizing comparable factors, the yield of the method of the invention is improved by over 100 times compared with a transfer bottle method in the prior art.
Description
technical field [0001] The present invention relates to the culture method of primary cell in the field of biological products, specifically a kind of utilization bioreactor and take polyester slice as carrier to cultivate primary animal cell (referring to in vitro culture no more than 5 generation cells after separating from tissue, hereinafter called primary cells). Background technique [0002] At present, there have been many reports on the use of bioreactors and microcarriers to cultivate animal cells at home and abroad. The cell microcarrier culture method was conceived and invented by Van Wezel in 1967. After 40 years of human exploration, this technology has become increasingly mature and perfect, and is widely used in the field of bioengineering and some cell products with important value. Microcarrier cell culture technology has many advantages that cannot be replaced by traditional cell culture technology. For example, in bioreactors, microcarriers increase the s...
Claims
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Inventor 王全新李杰
Owner LIAONING ANDI BIO TECH