Hello World,
I have been working with LS174t.
Complete Growth Media- 10% FBS/EMEM.
ATCC says to freeze with 5% DMSO/CGM.
when I defrost my vials, they are not attaching to flask.
vials put into Mr. Frosty filled with RT Isopropanol then put into -80 freezer for a day before being transferred to liquid nitrogen.
Do these cells freeze better with 10% DMSO?
or is there something that I am not aware of that is causing this freeze thaw trouble.
Thank you.
Hello Beatriz
LS174t cells may take 6-7 days to attach to the substratum. These cells grow in the form of islands and pile over one another. Subsequently they are released in suspension and they produce a lot of debris.
Regards.
Malcolm Nobre Thank you!
would you suggest to follow the ATCC protocol of changing media day 2/3.
Or aspirating, spinning down the suspended cells, then combining those with new media into the already attached cells.
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