Hybridoma Cell Culture Method
A technology of hybridoma cells and culture methods, which is applied in the field of cell culture, and can solve the problems of difficult enlargement and culture, complex structure, and difficult sealing
Active Publication Date: 2017-09-29
SHENZHEN INST OF ADVANCED TECH
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Problems solved by technology
[0004] Traditional microcarrier hybridoma cell culture reactors are mostly focused on the research of mechanically stirred reactors. This type of reactor has mechanical shear force, complex structure, difficult to seal, and difficult to scale up and cultivate; moreover, direct contact between air bubbles and cells is difficult. Facilitate the adherent growth of cells
The cell density of hybridoma cells during the culture process is not ideal, which limits the application of hybridoma cells in the large-scale industrial production of recombinant protein therapeutics
Method used
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[0021] The method for culturing hybridoma cells will be further described below in conjunction with the accompanying drawings and specific examples.
[0022] Such as figure 1 Shown, the culture method of the hybridoma cell of one embodiment, comprises the steps:
[0023] Step 110, inoculating the hybridoma cell suspension into a stirred flask containing macroporous microcarriers and medium and culturing under stirring conditions to obtain hybridoma cells grown on macroporous microcarriers, wherein the culture conditions are: culturing The pH of the base is 7.0~7.2, the mass fraction of serum in the medium is 5%, the culture temperature is 37°C, the mass fraction of dissolved oxygen is 30%, and the CO 2 The mass fraction of the medium is 5%, the concentration of the macroporous microcarrier in the medium is 1.2-1.5 g / L, and the stirring rate is 20-40 rpm.
[0024] In step 120, the hybridoma cells grown on the macroporous microcarrier are gradually enlarged and cultivated in a...
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The invention relates to a method for culturing hybridoma cells, comprising the following steps: inoculating the hybridoma cell suspension into a stirring bottle containing macroporous microcarriers and culture medium and cultivating them under stirring conditions to obtain microcarriers grown on macroporous microcarriers Hybridoma cells grown on macroporous microcarriers were scaled up and cultured in a stirred flask step by step, and at the same time, the serum content in the medium was gradually reduced until it was zero, and hybridoma cells adapted to serum-free medium were obtained ; Adopting continuous perfusion culture, the culture medium is a serum-free growth medium, and the hybridoma cells adapted to the serum-free medium with a density greater than 1×107 cells / mL are obtained, that is, hybridoma cells with high cell density.
Description
technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for culturing hybridoma cells. Background technique [0002] Animal cells can produce many important biological products, and large-scale animal cell culture is an effective way to produce recombinant protein therapeutics. Therefore, large-scale animal cell culture technology has become one of the most important key technologies in the field of biopharmaceuticals. [0003] The main method of traditional industrial production of recombinant protein therapeutic drugs expressed by animal cells is to use various animal cell culture reactors to culture hybridoma cells, insect cells, microorganisms, etc. Recombinant protein products. Among them, high-density culture of animal cells is the key to efficiently obtain a large amount of recombinant protein therapeutic drugs. [0004] Traditional microcarrier hybridoma cell culture reactors are mostly focused on the research ...
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IPC IPC(8): C12N5/02C12N5/20
Inventor 万晓春党利君金言
Owner SHENZHEN INST OF ADVANCED TECH