Novel serum-free culture medium
A serum-free medium, protein-free medium technology, applied in vertebrate cells, artificial cell constructs, animal cells, etc., can solve the problems of unclear, affecting the culture effect, and forming agglomeration.
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Embodiment 1
[0029] Embodiment 1: preparation of serum-free medium
[0030] The preferred components and dosage of the serum-free medium dry powder suitable for large-scale animal cell culture of the present invention are as follows:
[0031] Medium 1:
[0032]
[0033]
[0034]
[0035]
[0036] Medium 2: No heparin component, other components and dosage are the same as medium 1
[0037] Medium dry powder configuration method:
[0038] Add 1L of dry powder into 900ml of ultrapure water at a temperature of about 35°C. After stirring for half an hour, add a certain amount of sodium hydroxide to aid dissolution, then add concentrated hydrochloric acid and sodium bicarbonate to adjust the pH to 6.5-7.5. Filter with 0.2um negative pressure, aliquot into 500ml glass bottles and store aseptically in the dark.
[0039] The physical and chemical property parameters, detection methods and detection results of the medium obtained in the present invention are shown in Table 1.
[0040...
Embodiment 2
[0043] Example 2: Comparison of cell culture effects of different culture media under the same culture conditions
[0044] The culture conditions are shown in Table 2 below:
[0045] Table 2: List of Cell Culture Conditions
[0046]
[0047] As in the above culture method, the inventors respectively packed 300ml of the above-mentioned prepared three kinds of medium into 1000ml shake flasks, then added CHO cells, and the inoculation density was 6.0×10 5 cells / ml, put CO2 in the incubator (CO 2 5%), the temperature was 37°C, 20 μl was sampled every 24 hours, the cells were counted by a cell counter (purchased from invitrogen), the cell survival rate was detected by the MTT method, and the cell clustering was observed by a light microscope. The experimental results are shown in Table 3:
[0048] Table 3: Test results of cell density, cell viability and cell clustering in different media
[0049]
[0050]
[0051] The above experimental results show that the medium 1 ...
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