Dear All,
I am currently growing a b-cell line on top of a layer of endothelial cells. Now I wish to obtain a pure population of endothelial cells to carry out functional studies. However, due to the stickiness of the B-cells, it was never possible for me to obtain a pure EC population. I have tried removing cells with excessive PBS washes, in doing so the endothelial monolayer will also be lifted. I've also tried trypsin EDTA as well as none-enzymatic cell dissociation buffer from sigma but both detach B-cells just as efficiently as my endothelial cells. How can I resolve this problem without FACSing? Thanks!!!
Hi Tommy,
You can do a positive selection on your B cells using magnetic cell separation to separate them from the negative fraction (endothelial cells) that would remain unlabelled. Biotin positive selection kits can be purchased from Miltenyi or StemCell and then you could pick whichever biotin labelled antibody you want to mark the B cells.
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