culturing problems

Stephen Dahl stayve-and-irayne at worldnet.att.net
Tue Oct 5 23:13:11 EST 1999


"Elizabeth Aluck" <ealuck at HOTMAIL.COM> wrote in message
news:19991005215720.33477.qmail at hotmail.com...
>We recently built a new cell culture
> laboratory and I have been having an unusually high number of culture
> problems.  About 15 different cell lines which I have successfully grown
in
> other labs are not growing in this new lab.  I have eliminated obvious
> problems, such as media, CO2%, and temp.

Not to be a pain, but can you define "not growing"?  Are we talking dead or
slow to divide?  Is there debris in the media?  Are the cells shrinking in
size suggesting apoptosis?  Given that you have tried 15 lines perhaps you
could post the history of one.  That would save a back-and-forth of 20
questions as we try to divine an answer :o)

I am curious about a few things though...is the new facility outfitted with
brand new incubators, paint, flooring, etc.?  Is this a new building or
rennovated space in an existing one--for example, do you have contractors
working the swing or graveyard shifts who might be shutting down your
electrical to do wiring work?  Also, were your cells frozen prior to the
move and won't recover when thawed or are you moving them from the old to
the new space?  Finally, if you could explain how you ruled out the media,
CO2, etc.  For example, is your media made in house or purchased?  When you
say you checked the CO2 do you mean with a fyrite (shoot is that how its
spelled?  It doesn't look right) or do you mean the CO2 tank from the old
lab moved into the new?  I assume you've checked for mycoplasm.

Give me a litel more info and I'll mull this over some more.

If you prefer not to post your dirty laundry then e-mail me at work at:
stebby at jhmi.edu

Regards,

Steve Dahl
Dept. of Medicine/Renal
Johns Hopkins University






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