Removing T-cells from B-cells

palmer at msvax.mssm.edu palmer at msvax.mssm.edu
Thu Nov 16 11:16:44 EST 1995


Hello-
	Firstly, I'd like to thank every one who gave me in put
on my last posting(a question about BrdU staining for Flow),
I recieved several very helpful and thoughtful responses.

	My current problem involves removing T-cells from B-
cell preps.  I have been doing primary B-cell culture
experiments on cells isolated from human blood.  I isolate
them with Ficoll and AET-SRBC rosetting.  This has been
working pretty well with 85% plus purity.

	Unfortunately, I have to culture them with various
cytokines for more than a week.  The contaminating T-cells
tend to take off like a rocket and quickly out compete the
B-cells and may be influincing the progress of the culture.

	My question is does any one have a simple, quick
protocol to remove these last few T-cells?  I would prefer
something that does not beat to heavily on my B-cell yeild.
and is suitable for between 5 and 20 million cells.

			Thanks in Advance,

			Richard Palmer

Palmer at MSVAX.MSSM.EDU
Box 1198
Mt. Sinai Med. Center
One Gustav Levy Place
New York, NY 10029

New York, NY 10029



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