ATCC Hs 405.Sk细胞

产品名称：ATCC Hs 405.Sk细胞
产品货号： CRL-7279
Species： human
Source/Application：skin
Morphology：
Growth Mode：
Monolayer subculturing
1、Bring the trypsin-EDTA solution , balanced salt solution [Dulbecco’s Phosphate Buffered Saline without calcium or magnesium, and complete growth medium to the appropriate temperature for the cell line. In most cases, this is the temperature used to grow the cells (usually 37°C). For some sensitive cells, the trypsin-EDTA solution may need to be used at room temperature or 4°C.
2、Remove and discard the cell culture medium from the flask.
Rinse the cell monolayer with Dulbecco’s PBS without calcium or magnesium and remove.
3、Add 2 mL to 3 mL of the trypsin-EDTA solution and incubate at the appropriate temperature. Check the progress of cell dissociation by microscopy. To avoid clumping, do not agitate the cells by hitting or shaking the flask while waiting for them to detach.
4、Once the cells appear to be detached (5 to 15 minutes for most cell lines; they will appear rounded and refractile under the microscope), add 6 to 8 mL of complete growth medium with a pipette to the cell suspension to inactivate the trypsin. Gently wash any remaining cells from the growth surface of the flask. Check the cells with the microscope to be sure that most (>95%) are single cells. If cell clusters are apparent, continue to disperse the cells with gentle pipetting.
5、Add 12 mL to 15 mL of fresh culture medium to a new flask and equilibrate this medium to the appropriate pH and temperature.
6、Count the cells in suspension and determine their viability or simply divide them according to a routine split ratio and dispense them into the medium of the newly prepared flask. Do not add a concentrated cell suspension to an empty culture vessel as this can result in uneven cell attachment and growth.
7、Place the flask back into the incubator. Examine the culture the following day to ensure the cells have reattached and are actively growing. Change the medium as needed; for most actively growing cultures two to three times per week is typical.