Wash cells OR just add medium?????

[sexychicky!]

just wondering... to spin down and wash cells after trypsinisation or just add medium to the cell / trypsin suspension. I personally have never had any problems by just inactivating the trypsin by adding fresh medium, but would like to hear your opinions too..

[admin]

Hey there chicky,

the easiest is to wash cells in medium. Spinning down may harm the cells un-necessarily...

[jpap]

Personally, I add media to the cell-trypsin suspension, spin and resuspend in fresh media. Trypsin alone would not harm the cells, but I always add EDTA to the trypsin stock, it helps detaching considerably, and if not removed afterwards, (especially if you split the suspension 1:3 or 1:4) it may prevent the cells from adhering again.
Of course, if your cell type is sensitive to centrifugation you should avoid that protocol

[kiki06]

Yes, good points here hope they helped you Chicky.

[aftabac]

Hi
yes good points. here in our lab we also add new medium directly to the cells after trypsinization and with our cell lines it does not harm the cells, we also check the protocol in which the cells after trypsination and adding media are centrifuged gently, but it has the same results, but it very much depends on the type of cell lines you use, so you must set the protocl according to your cell lines, some suits one and other suites the othe one

regards
aftab

[molecule2005]

Yes I agree with Aftab,

you must check how your cells do with either method - centrifugation and directly adding media.

I would go for directly adding media as its easier and less stress to cells

[lolo]

good information......very nice.....

[sexychicky!]

thanx guys!!