cells in culture

WS via methods%40net.bio.net (by novalidaddress At nurfuerspam.de)
Thu Jan 25 18:12:58 EST 2007


Dear Pow,

you have may compassion. I took mine out from liquid nitrogen after
freezing them over the holidays and they simply refused to grow, they
look terrible indeed. Same for BOSC23 (a 293 derivative). 3T3 mouse
fibroblasts and H4 rat hepatoma in contrast grow more than well.I
treated all of them the same way before christmas (trypsinize, spn
down, resuspend, freeze), used the same freezing medium (90% FCS, 10%
DMSO) and resuspended all of them in DMEM high glucose, 5% FCS wis P/S
as usual. 293 HEK should grow in almost everything, actually.

Must be either the moon phase, not giving them christmas presents or
reading them good night fairy tales or simply a common cold (as I just
had one and 293 are humans, too). Couldn't hear them cry or sneeze yet,
however.

Tried to wash away the dead ones, trypsinized them and gave them fresh
food. Will have a look tomorrow and maybe take a new batch out of the
cold if necessary.

Maybe you have some conditioned medium handy (S/N of almost any healthy
confluent cells, passed through a .2 or .45microns filter), that
sometimes works, too.

Good luck!

Wolfgang

On Jan 25, 12:04 am, "Pow Joshi" <pow.jo... At gmail.com> wrote:
> Hi Folks,
>
> this is sheer desperation: I was wondering if anyone would have any
> idea why some cells (in this case HEK) are having problems attaching
> to the culture plate, and then die a sad horrible death.... all
> possible explanations are welcome including: "Pow, you've lost your
> touch at Tissue culture, even if you've been doing it for half your
> life". Thanks a bunch
> 
> pow



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