Protein-free serum-free medium suitable for diploid cell culture and its application
A technology of serum-free medium and diploid cells, which is applied in the field of cell engineering, can solve the problems of increasing the risk of culture, unfavorable product purification, etc., 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
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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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