culture cells and human plasma

Esteban Gabriel Vega egvega at hotpop.com
Sun Mar 20 23:55:14 EST 2005


"Dr Engelbert Buxbaum" <engelbert_buxbaum at hotmail.com> escribió en el 
mensaje news:d1bip0$vd0$01$1 at news.t-online.com...
> 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.

I agree with you Dr. and i'd like to add the following:
if you Harald want to work with plasma, try another anticoagulants such as 
heparin
which uses a diferent mecanism to prevent clotting.

EGVega 





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