serum free MDCK

Frauke Naumann fnaumann at scan.genetik.uni-koeln.de
Fri Sep 18 14:59:31 EST 1998


William Sun schrieb in Nachricht <6tmlbi$kvo at neuro.usc.edu>...
>Hello all,
>
>I purchased from ATCC an MDCK cell line adapted to serum free conditions.
To
>my dismay, I find these cells very difficult to detach from the dish with
>trypsin.  Does anyone out there share this problem?  The small percentage
>that gets removed and subcultured has low viability.  There is always many
>dead cells floating around.  The tech support at ATCC is of little help.
>Any comment/suggestions are appreciated.
>
>--
>William Sun, Ph.D
>Metabasis Therapeutics, Inc.           Phone: (619) 622-3930
>9390 Towne Centre Drive                FAX:   (619) 622-5545
>San Diego, CA 92121                    Email: william at alum.mit.edu

Hi William,
although "my" MDCK were not adapted to serum free conditions, detachment
took them up to 5 min at 37°C in 0,25% trypsin (I watched that through a
phase contrast microscope). And they were dividing just happily ever after.
If you did not take enough cells, perhaps they were "feeling lonely" and
were thus not viable? Passaging too few cells was always critical in my
experience.
Good luck
Frauke









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