TC Update
I don't want to jinx anything, but I have been thawing cells one line at a time and they are - so far - growing pretty well. But I am still a little verklempt because I think multiple problems were occurring simultaneously. And we all know it's shit having more than one variable at a time!
I ordered an expensive new cells from a company a couple weeks ago, thawed them out in the fresh and clean incubator (with no other occupants) and guess what?
Yep. Not the bleb monsters, but the spindly, leaving bits of themselves behind, non-mitotic problem again.
Turns out we (the lab) had switched FBS (fetal bovine serum) from HyClone to Atlanta Biologicals because we "got a good deal" and "the 3T3 cells looked okay". Gah! (By the way, the 3T3's don't really look okay.) So I threw some Clonetics FBS on my cells (along with some mycoplasma treatment, just in case) and lo and behold: they expand, with reasonably normal looking morphology.
My peoples - you can't just go around changing FBS and expect everything to be hunky-dory.
I think the terrible problems of April-May were a combination of cells being sickly, not liking the discount FBS (expensive tastes, wouldn't you know), and being consequently even more sensitive to whatever maux was griming up the incubators and t.c. hoods.
So now: lots of experiments, and ordering expensive things, and more experiments. And writing a paper. I really need to get this story done and out.
I ordered an expensive new cells from a company a couple weeks ago, thawed them out in the fresh and clean incubator (with no other occupants) and guess what?
Yep. Not the bleb monsters, but the spindly, leaving bits of themselves behind, non-mitotic problem again.
Turns out we (the lab) had switched FBS (fetal bovine serum) from HyClone to Atlanta Biologicals because we "got a good deal" and "the 3T3 cells looked okay". Gah! (By the way, the 3T3's don't really look okay.) So I threw some Clonetics FBS on my cells (along with some mycoplasma treatment, just in case) and lo and behold: they expand, with reasonably normal looking morphology.
My peoples - you can't just go around changing FBS and expect everything to be hunky-dory.
I think the terrible problems of April-May were a combination of cells being sickly, not liking the discount FBS (expensive tastes, wouldn't you know), and being consequently even more sensitive to whatever maux was griming up the incubators and t.c. hoods.
So now: lots of experiments, and ordering expensive things, and more experiments. And writing a paper. I really need to get this story done and out.
posted by Joolya at
12:05 AM
2 Comments:
We had cells.
We thawed them.
(Our -70 defroze itself the day afterwards, but that is another story and as all my cytokines were elsefreezed, someone larfed like a drain, and I heard it. Wun't me, though, of course.)
Anyway, we had cells an' we thawed them.
An' lo, a miracle did occur!
A few billion years of evolution did unroll themselves thenwards, as we all stood amazed...
For, lo, our cells did have flagellae, an' did swim mos' marvellous quick...
Where have you gone, Joolya? We miss your posts. Perhaps you could blog about physics and explain the wonders of the universe to us?
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