Shanghai Huyu has long-term supply of ATCC cell strains, with a complete range of products to ensure quality, if necessary, contact the company for sales. The basic technique of semi-adherent cell passage.
1. Cells with good cell growth status (90-95%) and cells with growth number greater than 5×105 were used for passage.
2. Use a pipette to thoroughly blow the wall of the culture dish to make the semi-adherent cells fall off. (If the cells are attached to the wall, digest it with trypsin for 1-3 minutes, and digest with 1 or 2 drops of serum).
Note: The ratio of digestive juice is: 0.25% trypsin + 0.02% EDTA. If the adherent cells gradually become round and partially suspended, the cells can be gently tapped with fingers, and most of the adherent cells are detached from the side or bottom of the flask, and then the cell stop solution is immediately added to terminate the cell digestion. The digestion time of each cell is different. Watch it when digesting. Do not digest for too long.
3. The mixed suspension was aspirated into a 15 ml centrifuge tube, centrifuged at 1000 rpm*3 min, and the original medium was removed.
4. Discard the supernatant from the centrifuge tube and resuspend the cells by adding the cell culture medium. The cell count was adjusted to 5 x 105 / ml.
5. After the cells are divided into bottles, add appropriate amount of cell culture medium to the cell culture flask, and place in a 5% CO2, 95% air, 37oC incubator for 24 hours.
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