A kind of culture method of cho cell expressing human follicle-stimulating hormone highly
A human follicle-stimulating hormone and high-efficiency expression technology, applied in the field of biopharmaceuticals, can solve the problem of low expression of human FSH, and achieve the effects of stable and controllable culture process, simple process flow and stable quality
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Embodiment 1
[0041] This embodiment provides a method for culturing CHO cells highly expressing human follicle-stimulating hormone, the steps are as follows:
[0042] After taking out the seed cells from the liquid nitrogen tank, melt them in a water bath at 37°C, take the cell suspension into a 15ml centrifuge tube, centrifuge at 1000rpm for 4min, and discard the supernatant. After resuspension with fresh basal medium preheated at 37°C, the basal medium consists of 18g / L CD-CHO medium, 0.2mM methionine, 0.3g / L sodium bicarbonate and 1.0g / L Composition of F68 of L. Cultivate in a 250ml air-permeable shake flask with a culture volume of 30ml, and the culture conditions are 120rpm rotation speed, 37°C temperature, 75% humidity, and 5% CO2. Seed expansion is passed step by step in the order of 250ml shake flask-500ml shake flask-1000ml shake flask-2000ml shake flask. After 7 days of expansion, the cell culture volume reaches 500ml, and the density is 3.0×10 6 pieces / ml.
[0043] Take the s...
Embodiment 2
[0046] This embodiment provides a method for culturing CHO cells highly expressing human follicle-stimulating hormone, the steps are as follows:
[0047] After taking out the seed cells from the liquid nitrogen tank, melt them in a water bath at 37°C, take the cell suspension into a 15ml centrifuge tube, centrifuge at 1000rpm for 4min, and discard the supernatant. After resuspension with fresh basal medium preheated at 37°C, the basal medium consists of 18g / L CD Opti CHO medium, 0.2mM methionine, 0.3g / L sodium bicarbonate and 1.0g / L Composition of F68 of L. Cultivate in a 250ml air-permeable shake flask with a culture volume of 30ml, and the culture conditions are 120rpm rotation speed, 37°C temperature, 75% humidity, and 5% CO2. Seed expansion is passed step by step in the order of 250ml shake flask-500ml shake flask-1000ml shake flask-2000ml shake flask. After 8 days of expansion, the cell culture volume reaches 500ml and the density is 3.5×10 6 pieces / ml.
[0048] Take t...
Embodiment 3
[0051] This embodiment provides a method for culturing CHO cells highly expressing human follicle-stimulating hormone, the steps are as follows:
[0052] After taking out the seed cells from the liquid nitrogen tank, melt them in a water bath at 37°C, take the cell suspension into a 15ml centrifuge tube, centrifuge at 1000rpm for 4min, and discard the supernatant. After resuspending with fresh basal medium preheated at 37°C, the basal medium consists of 20g / L Ex-cell 302 medium, 0.2mM methionine, 0.3g / L sodium bicarbonate and 1.0g / L F68 composition. Cultivate in a 250ml air-permeable shake flask with a culture volume of 30ml, and the culture conditions are 120rpm rotation speed, 37°C temperature, 75% humidity, and 5% CO2. Seed expansion is passed step by step in the order of 250ml shake flask-500ml shake flask-1000ml shake flask-2000ml shake flask. After 8 days of expansion, the cell culture volume reaches 500ml and the density is 3.1×10 6 pieces / ml.
[0053] Take the seed...
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