Isolation of monocytes
bkournik at dres.dnd.ca
Thu Jun 30 14:52:05 EST 1994
In article <92940628192029.0004500605PK1EM at mcimail.com> JOHNSON+c_MICHAEL%A1%Mallinckrodt at mcimail.com ("JOHNSON, MICHAEL") writes:
>From: JOHNSON+c_MICHAEL%A1%Mallinckrodt at mcimail.com ("JOHNSON, MICHAEL")
>Subject: Isolation of monocytes
>Date: 28 Jun 1994 12:36:41 -0700
>I need to isolate monocytes from a large amount (@ 2 l) of porcine blood. I
>would like to know if anyone is doing work of this nature and what method would
>be the most efficient. Also, what medium is required for culturing these cells?
I have isolated mononuclear cells from porcine blood using Histopaque 1077.
Blood was collected using Acid Citrate Dextrose Sol'n B as an anticoagulant
(Heparin and EDTA did not work well) and the cells were isolated by layering 6
mL blood over 3 mL Histopaque in a 15 mL tube and centrifuging for 30 min at
800 x g. Clearly this isn't too practical for a 2L volume but you may be able
to scale the process up a bit by using a larger test tube. Do you really need
to process 2L at once or could the processing be spread out over time?
Perhaps if you re-posted with more detail of what you'd like to accomplish you
might get some additional suggestions from the net.
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