Human bone marrow processing using Cobe 2991 and CS 3000 blood cell separators for further ex vivo manipulation

Several automated procedures are now available to enrich stem cells from large bone marrow (BM) volumes prior to ex vivo treatment or cryopreservation. This report details our experience using a ficoll-hypaque (F/H) gradient on Cobe 2991 cell washer and a CS 3000 continuous flow separator, on 90 BM processed for allogeneic and autologous transplantation. In the Cobe series, from 70 BM aspirates, 89 +/- 5% of the original mononuclear cells (MNC) was found in the light density fraction with a erythrocytes (RBC) and granulocytes (PMN) removal of 98 +/- 1 and 97 +/- 4.5%, respectively. Over 80% of the initial myeloid precursors (CFU-GM) were recovered in a small final volume. Twenty BM processing were performed with a CS 3000 separator using program "3" and granulo chamber. This technique yielded 86 +/- 9% of the initial MNC while 85 +/- 10% of RBC and 90 +/- 1.1% of PMN was removed. Over 75% of the original CFU-GM was recovered in the final product. Both techniques are effective to large-scale purification of progenitor cells and readily available as routine procedures for marrow processing.