Protocol Online logo
Top : Forum Archives: : General Lab Techniques

Do you reuse cell culture vessels? - (Apr/11/2005 )

If I run short of cell culture vessels I sometimes reuse them such as after trypinizing cells in a flask I replant the cells into the same flask. Is that OK? If you do the same, how many times a vessel can be reused?

thank you for your input.


I have not done this but was recently asked about it. My concern would be that the bottom of the flask will be coated with intracellular matrix and secreted proteins that may influence the growth of your cells. It may be alright depending on what you are using the cells for?


HiYa PostDoc,

for many cell lines, when maintaining the stock culture, reusing the flask is not a problem ... I have had no problem with adherent cells like: CHO, Cos-7, DU145, HeLa, and some neuroblastoma lines and of course suspension cells are okay: CEM and Molt-4 are the ones I have mostly used. I use flasks at least twice, only going for a third run if keeping the stock as a backup (i.e. may require extra cells).

I have noticed two cell lines that won't grow well in the flask previously used to culture the stock and these were U251 and U87 (glioma cell lines).

If I am setting up cells for an experiment, then I generally use a new flask.

So ... for your cells, try it smile.gif. It does save money and all bosses would be happy with that rolleyes.gif

Good Luck



I completely agree with AussieUSA's comments. In our lab we reuse the T-flasks for passaging fairly continuously - sometimes greater than 5 passages sometimes 10 in the same flask. With some cell lines it seems to help as they prefer their matrix rather than plain plastic to adhere to. This has never given us any problems. However, like AussieUSA said, any experiments are always done in new flasks or dishes.