Separation of Mononuclear Bone Marrow Cells using the Cobe 2997 Blood Cell Separator

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Abstract
In the present study, we report the results of our evaluation of the use of the continuous‐flow cell separator Cobe 2997 to isolate from human bone marrow (BM) aspirates the mononuclear cell (MNC) fraction containing hematopoietic stem cells. This MNC concentrate is isolated in 15% of the original BM volume and contains 23% of the initial nucleated cells. It is enriched as concerns the BM MNC fraction (lymphocytes + monocytes recovery; 80%), whereas the contamination with granulocytes, red blood cells and platelets is reduced to 7.2, 1.5% and 41%, respectively, of the cells initially present in the BM suspensions. Furthermore, it is demonstrated that this MNC concentrate is highly enriched in granulocyte‐macrophage‐colony‐forming cells (CFU‐GM; recovery 83%). The method is simple, inexpensive, efficient and reproducible. It allows rapid processing of a large volume of BM without substantial loss of hematopoietic progenitor cells. It represents a valuable method of BM MNC concentration prior to further in vitro manipulations such as T cell or tumor cell depletion or cryopreservation.