culture cells and human plasma
Dr Engelbert Buxbaum
engelbert_buxbaum at hotmail.com
Thu Mar 17 04:31:09 EST 2005
Harald Heider wrote:
> I am trying to add human plasma (derived from citrate blood) to cell
> cultures. The set-up is the following: HEK293 cells are cultured in 12
> well-plates in 1 ml of complete medium (DMEM with serum and
> antibiotics). Then 700 microliters of human plasma are added (without
> removing the medium). After 4h the whole stuff is coagulated! It is not
> the medium and not the incubation period (I tried this). Does anybody
> know how to avoid the clotting under these circumstances?
Apparently the citrate is not sufficient to prevent clotting under these
circumstances, after all the medium will contain Ca and Mg to keep the
cells alive. Your best chance is probably to convert your plasma to
serum by adding some Ca and incubating until coagulation is complete.
You may have to centrifuge. If you perform the incubation at 56 degrees
C you heat-inactivate the complement as well, this may also reduce the
risk of viral infections.
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