Isolation of monocytes

Bill Kournikakis 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

>Netters,
>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?

>Thanks,
>Michael Johnson

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. 




Bill Kournikakis
Medical Countermeasures Section
Defence Research Establishment Suffield
Medicine Hat, Alberta, Canada  

If research was easy we'd call it "Search"
	



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