Comparison of Progenitor Cell Concentration Techniques: Continuous Flow Separation versus Density-Gradient Isolation

Bone marrow harvests were processed using two techniques; 20 mononuclear cell concentrations (MNC) were prepared on the COBE Spectra and compared to 10 light-density cell fractions isolated using the COBE 2991. Both procedures recovered essentially the same percentage of nucleated cells (22 versus 21%) and gave progenitor cell recoveries of 132 and 100% of the CFU-GM and 101 and 104% of the CD34⁺ cells in the MNC and light-density products, respectively. The advantage of the Spectra MNC concentrate is that it was prepared with reagents approved for injection by the Food and Drug Administration. However, the average hematocrit on the MNC concentrate was 4%, while it was unmeasurable in the light-density cell fractions. This difference is significant only when bone marrows require purging with 4-hydroperoxycyclophosphamide or erythrocytes otherwise interfere with processing. The Spectra procedure recovered a larger percentage of MNC cells and had less contamination with mature granulocytes than did the density-gradient technique. When erythrocytes do not affect the bone marrow processing protocol, a Spectra MNC concentrate is a safe substitute for a light-density cell preparation.