change adherent cells into suspenion cells - (Feb/24/2006 )
I would like to know if anyone knows how to make an adherent cell line into a suspension cell line.
If you could point me in the right direction please advise.
We are using CHO "Chinese Hamster Ovary"cells to patch clamp. The protocol we are using is to use an enzyme to digest the cells membrane to get them to detach off the bottom of a T-75 flask. We would like to eliminate this step and have the cells transformed into a suspension cell line. This step we are currently using to remove the cells from the flask is making the membrane too fragile.
Thank You in advance.
Do you know if your cells can be made into a suspension culture?
Generally, suspension cultures grow in spinner flasks at 60-80rpm and the addition of heparin to the media helps prevent the cells from aggregating.
You could also try sloughing to get the cells off the flask. I work with High Five cells (moth ovarian cells) and use this to get them off. Sloughing involves removing all but 5ml of the media, standing the flask upright, sucking up a small amount of the remaining media into a pasteur pipet and then essentially blasting the cells off the surface. It can be done quickly and if done right will only kill 1% of the cells. It takes a bit of knack though.