Handling Neuronal Stem Cell Cultures in a Cell Production Facility
筆記録
Place three Petri dishes in the water pan at the base of each incubator. Fill these dishes with sterile water, and maintain the water level in these dishes as they are critical for maintaining the relative humidity set point in the incubator.
Within the graphical interface, click on Incubator. Subsequently, click on the existing value for relative humidity under the set point and enter 85%. Then, click on the green check mark to accept this value.
Next, adjust the buffer chambers, the process chamber, and an incubator to the gas set points required for the type of cells being introduced. Clean the surface of the process chamber with sterile gauze, and a non-flammable disinfectant, such as benzalkonium chloride-based products.
Then, clean the gloves and surfaces that are commonly touched, such as door handles. Subsequently, clean the surface of the laminar flow hood and buffer chamber with 70% ethanol and allow it to dry.
After that, place the flask or plate of cells in the hood, and briefly wipe its exterior surface with a sterile gauze sponge saturated with ethanol. Then, put the flask or plate in the buffer chamber and run a dilution factor. Upon completion, immediately transfer the flask or plate to the appropriate incubator.
As the incubator is at the back of the process chamber, pull a shelf out into the process chamber for cell placement and avoid opening the incubator doors unnecessarily or for extended periods of time.
In this step, prepare the cell culture medium. Allow the medium to equilibrate with the carbon dioxide and oxygen levels present within the system by leaving the media container slightly uncapped for 20 minutes.
Afterward, remove the old medium from the wells. For pluripotent stem cells, remove almost all of the old medium with a small amount of the old medium remaining, to prevent desiccation during the feeding process. For neural stem cells, remove half of the old medium. Then, add fresh medium to the cell culture plates, and place the plates back into their designated incubator.
