Re-using Culture flasks - (Jun/01/2009 )
Hi dear, you can use culture flask 4-5 times unless you devoid trypsin from it. Either wash the flask using PBS or Culture medium, then add cell 
s.
how do you devoid trysin?
pbs wont get rid of previous inhabitants, nor will culture media
not sure what your on about rock
Stephan on Jun 1 2009, 05:16 PM said:
Does anybody else do this on a regular basis and any other thoughts?
ya,we regularly reuse culture dishes upto 3 times.....there is no problem....but rinse properly
If I wash the plate well with PBS, can I reuse 6 well plates?
THE ROCK on Jul 10 2009, 06:59 AM said:
s.HMG on Sep 9 2009, 09:46 AM said:
Buddy on Dec 6 2009, 04:53 PM said:
If you all read the above posts you will see that you are in error... re-using is a problem, if you do it you are jeopardizing you results through a number of factors including cellular damage, cell type drift (lab evolution), and general contamination issues.
Be better scientists!
I would like to convince my boss, that it is a bad idea to re-use flasks. Can anybody recommend some litterature regarding this topic?
Thanks
We can re-use flask depending on our cells viability and confluencity...... But i would not re-use flask more than 2 times... If not, i would do a new batch of cells 
we do re-use flask for the sake of saving cost
but have to be reasonable...
for adherent cell, 1-3 time is still ok ( despire the contamination problem, as you re-use more time, the adhesive protein may build up on the plastic surface and will affect the cell growth/metabolism.... the most obvious evidences is u will find very hard to trysinise cell in the late)
for suspension cell, no prob, at least in our hand (more than 10 time). but yet, u will notice some dirt-like residue on the plastic after few time re-use(which i think it might be the protein residue...)
in conclusion, be moderate.
ask yourselves, save cost or better science
tongck on Jun 3 2010, 06:38 AM said:
but have to be reasonable...
for adherent cell, 1-3 time is still ok ( despire the contamination problem, as you re-use more time, the adhesive protein may build up on the plastic surface and will affect the cell growth/metabolism.... the most obvious evidences is u will find very hard to trysinise cell in the late)
for suspension cell, no prob, at least in our hand (more than 10 time). but yet, u will notice some dirt-like residue on the plastic after few time re-use(which i think it might be the protein residue...)
in conclusion, be moderate.
ask yourselves, save cost or better science
Please listen to bob1.....he as usual is 100% correct and is posting on here to give best advice. From some of the comments posted so far, he is wasting his time.
Kindest regards
Uncle Rhombus.
tongck on Jun 2 2010, 10:38 PM said:
but have to be reasonable...
for adherent cell, 1-3 time is still ok ...
my experience with adherent cells is that you can almost never get them all off from the edges of the flask... i tried re-using the flasks when i first got my adherent cells, and i quickly noticed that under the microscope the edges were 100% confluent while the rest was just barely seeded.
just one or two cells start apoptose from being packed in so tight and there's your background level of inflammation when the assays are finally done
*edit* also, the crap leftover from the previous culture (fragments of protein, dna etc) in my mind is pretty much impossible to remove except by scrubbing or with a water jet... if you have ever noticed clumping after trypsinization it can be due to dna strands, cells with human dna sticking to their outside can be a cause background inflammation as well