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Dear all,
During a recent PK study we discovered that bone marrow samples
(washed out from femur with PBS), if stored .at. -20*C, forms a gel-like
substance if thawed. The substance appeared to be a fiber/fat/protein
"ball" and will absorb some of the drugs. This happens with various
mouse strains.
If anybody has a good way to store mice bone marrow samples for short
period of time (preferably frozen), could you please kindly post the
method?
Thanks a lot.
Sherwin
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Dear Sherwin,
How is the bone marrow thawed? It seems important for plasma(see
below), may be applicable to marrow.
Martin
: Transfusion. 1989 Feb;29(2):165-9. Links
Formation of a cryogel during processing of cell-free plasma.
Rock G, Palmer D, Tittley P, McCombie N, Schoendorfer D, Drago J.
Department of Medicine, University of Ottawa, Ontario, Canada.
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Hi Martin,
Thank you very much for your suggestion.
We thawed the frozen bone marrow samples the same way we thaw the
plasma: room temperature for 1 hour. While we did not observe any
cryogel-like substance in cell-free plasma, we had that happening in
bone marrow homogenate (needle passage) every single time.
I do not have access to the full paper you quoted. After reading the
abstract, I think the "cell-free" part does not apply to our samples,
but the "slow-thaw at 4*C" might help. Will give it a try.
Any other comment/suggestion will be deeply appreciated. Thank you.
Sherwin
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Sherwin
Try thawing for 3h at RT with increasing conc. of DNase I. I hope when
you thaw you add media to cell and not vice versa...
Deepak
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Hi Deepak,
The sample we are harvesting is directly flushed out from mice femurs.
Therefore we have to use buffer to "wash" them out of the bone. Are
you suggesting the gel-like substance is DNAs precipitate out?
Thanks for the input.
Sherwin
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Hi Sherwin,
I am not sure about the question. Do you see the gel like substance
after you flush or when you thaw your samples?
Deepak
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Hi Deepak:
The gel-like substance only showed up after freeze-thaw cycle. The
fridge stored sample did not have this problem.
Sherwin
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