24 replies to this topic

[Felina]

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

Edited by Felina, 13 January 2010 - 11:17 AM.

[arera]

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  

[tongck]

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

[rhombus]

tongck, on Jun 3 2010, 06:38 AM, said:

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

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.

[telamon]

tongck, on Jun 2 2010, 10:38 PM, said:

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 ...

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

Edited by telamon, 04 June 2010 - 10:01 AM.

[Genecks]

I wonder how practical it would be to make an autoclavable culture flask.
I could imagine it:

Make a culture flask out of glass, like a general fischerbrand 75 cm flask. Have a cap and filter that can be autoclaved.

After use, wash it out really well, rinse with a good amount of ethanol, place it in the fumehood to dry, place the autoclavable cap on, and then bring to an autoclave to sterilize. Afterward, bring it to the hood, open it up, and use it.

Why is something like that not possible?

What about cell plates? i think it would be more practical to have cell plates. Use them, wash them, rinse them, wrap them with aluminum foil, and sterilize. Afterward, use in hood.

It seems like the major concern would be trace chemicals left behind rather than intruding prokaryotic or eukaryotic organisms.

It seems like a person might as well use a glass petri dish if he/she wants a reusable container. I see the only problem in being careful so no contamination occurs. Otherwise, yeah, I think some glass petri dishes wouldn't be such a bad idea.

Edited by Genecks, 05 May 2012 - 05:16 PM.

[leelee]

Genecks, on 05 May 2012 - 05:05 PM, said:

I wonder how practical it would be to make an autoclavable culture flask.
I could imagine it:

Make a culture flask out of glass, like a general fischerbrand 75 cm flask. Have a cap and filter that can be autoclaved.

After use, wash it out really well, rinse with a good amount of ethanol, place it in the fumehood to dry, place the autoclavable cap on, and then bring to an autoclave to sterilize. Afterward, bring it to the hood, open it up, and use it.

Why is something like that not possible?

It is possible. Before disposable plasticware, all tissue culture ware was glass!

But what a pain to have to decontaminate, then wash thoroughly, then sterilise every time you use a flask or dish? The time you waste doing that would buy you a case of disposables easily. The most valuable resource in any lab is your time.

Not to mention the stringent quality control you would need in order to maintain your glassware. Lab water supplies can (and do) have significant levels of endotoxin present in them too, so I don't know about you but I'd rather not have LPS in my tissue culture thanks!

Edited by leelee, 05 May 2012 - 06:58 PM.

[Genecks]

Well, right now I'm out of flasks. The lab tech didn't order more, and I'd jump for some glass petri dishes. lol.

[s1moninha]

Maybe trypsin is not the better option to dettach your cell from the bottle, see if you can use cell scrapers, they're much better opption when you can't dettach your cells using trypsin solution. Ad try to not reuse your bottles!!!

[kayjay21]

Will cell scrapers get off all the cells though? And couldn't they be damaged as a result of the scraping?

Edited by kayjay21, 17 May 2012 - 08:49 AM.