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change media everyday (or every other day) for MTT assay?


Best Answer antitoxin, 29 July 2013 - 08:31 AM

It depends on the estimated number of cells in your culture is the most commonly replaced every 48 hours should be made Otherwise  they are stressed

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#1 crom80

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Posted 25 July 2013 - 07:45 AM

I am performing an MTT assay and was wondering how others are letting cells grow for long term MTT assays (5-7day period). I seed cells and collect data 0, 1, 2, 3, 5 and 7 days after seeding.
I am currently adding additional media on day 3 for day 5 and 7 period wells as I noticed media evaporating.
Do people usually leave media as is or change everyday or every other day?

Thank you.

#2 aimikins

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Posted 27 July 2013 - 11:11 AM

well, you're using a treatment and at the end you're using MTT to measure cytotoxicity of your treatment, right? (this is what I've used it for)

 

I replace the media on the same schedule as I always would for that cell type. if it was a one-time treatment, just use regular media. if it's a continuous treatment, just make sure your drug is in the fresh media.

 

does that help? 


"it is a miracle that curiosity survives formal education" -A.E.

#3 antitoxin

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Posted 29 July 2013 - 08:31 AM   Best Answer

It depends on the estimated number of cells in your culture is the most commonly replaced every 48 hours should be made Otherwise  they are stressed



#4 crom80

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Posted 29 July 2013 - 12:23 PM

well, you're using a treatment and at the end you're using MTT to measure cytotoxicity of your treatment, right? (this is what I've used it for)

 

I replace the media on the same schedule as I always would for that cell type. if it was a one-time treatment, just use regular media. if it's a continuous treatment, just make sure your drug is in the fresh media.

 

does that help? 

 

I am not doing any treatments. I am comparing wild type and Protein of interest over expressed cell growth over time.

My main concern is the evaporation of the wells at the edges and was wondering if I replace media everyday would evaporation be less.

It depends on the estimated number of cells in your culture is the most commonly replaced every 48 hours should be made Otherwise  they are stressed

 

The cells in normal culture is given fresh media twice a week (monday and friday) so I didn't think about giving fresh media every 48 hours.



#5 aimikins

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Posted 29 July 2013 - 12:29 PM

oh, I'm sorry, I don't think we're talking about the same assay. I apologize for any confusion.


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