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Protein-free serum-free medium suitable for diploid cell culture and application

A technology of serum-free medium and diploid cells, which is applied in the field of cell engineering, can solve the problems of unfavorable product purification and increase the risk of culture, and achieve the goal of reducing the risk of allergic reactions, reducing the difficulty and cost of separation and purification, and simplifying the purification steps Effect

Active Publication Date: 2016-04-06
SICHUAN BAINUOJI TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] From the above analysis, it can be seen that there are at least two problems in the current production process. On the one hand, serum needs to be added to the medium; on the other hand, the medium contains unclear components such as protein, which increases the risk of cultivation. It is also not conducive to product purification

Method used

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  • Protein-free serum-free medium suitable for diploid cell culture and application
  • Protein-free serum-free medium suitable for diploid cell culture and application
  • Protein-free serum-free medium suitable for diploid cell culture and application

Examples

Experimental program
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Effect test

Embodiment 1

[0039] What this embodiment prepares is the culture medium that is suitable for MRC-5 cell culture, and the data that each component dosage can refer to the data given in Table 3, concrete steps are as follows:

[0040]Add Tween 80 into a reaction vessel with a capacity of 1000ml, then add 50ml of water for injection to dissolve, then add linoleic acid, linolenic acid, arachidonic acid, vitamin E, myristic acid, and stearic acid in turn, and wait until they are fully dissolved , rehydration with water for injection to 500ml, then sequentially add sodium chloride, glucose, fructose, stir until clarification, rehydration with water for injection to 900ml, finally add calcium chloride, magnesium sulfate, potassium chloride, disodium hydrogen phosphate, phosphoric acid Potassium dihydrogen, L-arginine hydrochloride, L-aspartic acid, L-asparagine, L-cystine dihydrochloride, L-histidine hydrochloride, L-leucine, L-isoleucine, glycine, L- Lysine hydrochloride, L-methionine, L-phenyla...

Embodiment 2

[0051] This example prepares a medium suitable for 2BS cell culture, and the dosage of each component can refer to the data given in Table 3, and the specific steps are the same as in Example 1.

[0052] In order to examine the performance of the medium provided in this example in the cell growth stage, 2BS cells were cultured with the medium provided in this example and MEM at a concentration of 10%. The specific implementation method adopted in this example is: take 2BS cells whose cell generation is 10-40 generations, and digest the monolayer cells from the culture flask with 0.05% trypsin and 0.02% EDTA (ethylenediaminetetraacetic acid) solution , resuspend the cells with the prepared culture medium of this example and MEM containing 10% serum concentration respectively, and all use 0.55*10 6 Inoculate cells / ml, add a certain amount of corresponding medium and culture at 37°C, digest and count every 24 hours.

[0053] In the following, the culture medium provided in this ...

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Abstract

The invention provides a protein-free serum-free medium suitable for diploid cell culture and a preparing method thereof. The medium contains calcium chloride, magnesium sulfate, potassium chloride, sodium dihydrogen phosphate, L-arginine hydrochloride, L-asparaginate, L-cystine dihydrochloride, L-histidine hydrochloride, L-leucine, L-isoleucine and the like. The medium can meet the requirement of the growth or maintenance stage of diploid cells and prevents contact with exogenous viruses possibly existing in protein and serum to reduce potential safety hazards; as the components are simple and definite, standard products can be purchased in the market, the difference between different product batches is small, and the step of purifying down-stream products is simplified; a scientific basis is provided for large-scale standard production. In addition, the protein-free serum-free medium can be applied in the growth or maintenance stage of the diploid cells.

Description

technical field [0001] The invention belongs to the technical field of cell engineering, and relates to a protein-free, hydrolyzate-free serum-free medium, in particular to a protein-free, hydrolyzate-free serum-free medium suitable for diploid cell culture and its Preparation methods and applications. Background technique [0002] Cell culture medium is the basic material for supplying cell nutrition and promoting cell proliferation when culturing cells in vitro. It is also the living environment for cell growth and reproduction. It is the basis of in vitro cell culture and is widely used in the fields of biological products and stem cell therapy. Protein-free, hydrolyzate-free serum-free medium is a kind of cell culture medium, its composition is completely defined, and it does not contain unknown substances such as protein and hydrolyzate. [0003] In the process of preparing vaccines with diploid cells, there are two stages that require the use of cell culture medium, n...

Claims

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Application Information

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IPC IPC(8): C12N5/07
Inventor 扶星星吴福文王永胜马海强刘丹
Owner SICHUAN BAINUOJI TECH CO LTD
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