Comparison of Two Methods for Concentrating CD34 + Cells from Patients with Acute Non-Lymphocytic Leukemia

Abstract

The aim of the present study was to compare two different methods for obtaining CD34 + cells from the peripheral blood or the bone marrow of patients with acute non-lymphocytic leukemia (ANLL). Twenty-two samples, obtained from 19 patients, were density cut (Ficoll-Hypaque 1.077) and, after incubation with My10 antibody, separated by panning or by immunomagnetic beads. Immunomagnetic beads provided a significantly better separation than panning, either in terms of concentration of CD34+ cells (85.5 ± 11.6% vs. 55.7±25.0%, p = 0.003) or in terms of depletion of CD34+ cells (3.9 ± 8.0% vs. 30.9 ± 26.3%, p = 0.008). This was consistent with the virtually complete depletion of colony forming cells (CFC) in the CD34 negative fraction and the recovery of virtually all the CFC in the positive fraction in the samples separated by immunomagnetic beads. In conclusion, separation by immunomagnetic beads can allow collection of nearly pure CD34+ and CD34—cell populations from patients with ANLL, thereby facilitating the study of the biological characteristics of these cell populations. Moreover the method is less time consuming than panning and is not toxic to the CFC.

Key Words:

• CD34+ cells
• acute non-lymphotic leukemia
• panning immunomagnetic beads