Hi,
I want to isolate hematopoietic stem and progenitor cells from human bone marrow that I get fresh. I only get small samples. So far I've been using Ficoll and then proceeded to staining and flow sorting afterwards. The sorted cells are then analyzed by NGS and FISH.
Since I'm utilizing the bone marrow right away for sorting of particular populations, would preparation of the bone marrow by just focusing on red cell removal increase my yield? All suggestions are welcome.
Peter
Hi, Peter.
You may try MACS cell separation. It's simple, no expensive equipment, fast and effective for the enrichment or isolation of specific cell separation with surface protein labelled. Just lyse RBCs, and critical to remove ECM from bone marrow-derived cells to be single cell suspension for MACS separation. Use milder trypsin-EDTA (1/5 dilution).
Best wishes.
What solution would you use to lyse RBCs and would this not be toxic to stem cells?
Best,
Peter
Here's commercial product although it can be made.
11814389001 Roche Red Blood Cell Lysis Buffer
It's not toxic at all to remaining cells.
Regards.
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