PBMC separation from brain tissue

Rick Arthur Bright rbright at emory.edu
Wed Mar 27 20:06:51 EST 2002


I am trying to analyze lymphocytes, monocytes, granulocytes, etc from
brain and lung tissues.  After collagenase digestion I, of course, have a
lot of junk...stuff I don't want to run through the instrument.

What is the best way of separating the cells from the junk?  I tried
Ficoll-paque, which nets me the lymphocytes, but I lose other cells types
that might be important.

Would a Percoll gradient be a better route to go?  What gradients would be
most appropriate.  I am digging in the literature as well, finding many
reference to percoll gradients, but few of them say what the gradients are
and where the cells of interest will settle.

Any quick thoughts would be helpful.  I, hopefully, will stumble upon that
magic reference sometime tonight, but if you know already I would
appreciate your help.

Thanks

Rick






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