Comparison of five methods for concentrating progenitor cells in human marrow transplantation

Enrichment of bone marrow (BM) aspirates is an important prerequisite prior to in vitro treatment or cryopreservation. In this regard, we have analyzed the results obtained on 190 BM processed by the following 5 techniques: HES sedimentation with centrifugation; COBE 2991 blood cell processor; Ficoll/hypaque (F/H) gradient centrifugation; Continuous flow cell separator (CS 3000 Fenwal); Semicontinuous blood cell separator (Dideco T 90). Each procedure was evaluated by measuring the recovery of nucleated marrow cells (NC), mononuclear cells (MNC), committed progenitor cells (CFU-GM), the reduction of BM volume and the removal of red blood cells (RBC) and polymorphonuclear cells (PMN). The results of this comparative study show that F/H gradient on a COBE 2991 cell washer provides the most efficient system for purifying a MNC fraction (89% recovery) from unwanted cells (RBC less than 2% and PMN less than 2%) in a very small volume (98% reduction) with a good recovery of CFU-GM (80%).