Relative frequency of immature CD34+/CD90+ subset in peripheral blood following mobilization correlates closely and inversely with the absolute count of harvested stem cells in multiple myeloma patients

Bela Balint; Ivan Stanojević; Milena Todorović; Dragana Stamatović; Mirjana Pavlović; Danilo Vojvodić

doi:10.2298/VSP170505090B

Bela Balint Institute for Transfusiology and Hemobiology, Military Medical Academy, Belgrade, Serbia; Institute for Medical Research, University of Belgrade, Serbia; Serbian Academy of Sciences and Arts; Faculty of Medicine of Military Medical Academy, University of Defence, Belgrade, Serbia;
Ivan Stanojević Military Medical Academy, Institute for Medical Research, Belgrade, Serbia The University of Defence, Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia
Milena Todorović Clinical Center of Serbia, Clinic for Hematology, Belgrade, Serbia
Dragana Stamatović Military Medical Academy, Clinic for Hematology, Belgrade, Serbia; University of Belgrade,The University of Defence, Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia
Mirjana Pavlović Department of Computer and Electrical Engineering and Computer Science, FAU, VL, USA
Danilo Vojvodić The University of Defence, Faculty of Medicine of the Military Medical Academy, Belgrade, Serbia Military Medical Academy, Institute for Medical Research, Belgrade, Serbia

DOI: https://doi.org/10.2298/VSP170505090B

Keywords: stem cells, hematopoietic stem cell transplantation, bone marrow, flow cytometry, multiple myeloma, antineoplastic combined chemotherapy protocols,

Abstract

Background/Aim. Stem cells (SCs) guarantee complete/long-term bone marrow (BM) repopulation after SC-transplants. The aim of the study was to evaluate absolute count of total SCs (determined by ISHAGE-sequential-gating protocol – SC^ish) and relative frequency of immature CD34⁺/CD90⁺ (CD90⁺SC^ish) subset in peripheral blood (PB) as predictive factors of mobilization and apheresis product (AP) quality. Methods. Mobilization included chemotherapy and granulocyte-growth-factor (G-CSF). Harvesting was performed by Spectra-Optia-IDL-system. The SCs^ish were determined as a constitutional part of CD34⁺ cells in the “stem-cell-region” using FC-500 flow-cytometer. In this study, the original ISHAGE-sequential-gating protocol was modified by introduction of anti-CD90-PE monoclonal-antibody into the analysis of CD90 expression on SC^ish (CD90⁺SC^ish). The results were presented as a percentage of SC^ish per nucleated-cell count, absolute SC^ish count in μL of the PB or the AP, percentage of the CD90⁺SC^ish expressed to SC^ish and absolute CD90⁺SC^ish count in μL of the PB or the AP. Results. The absolute count of total SC^ish and CD90⁺SC^ish was significantly higher (p = 0.0007 and p = 0.0266, respectively) in the AP than in the PB samples. The CD90⁺SC^ish/total SC^ish indexes from PB were higher than indexes from the AP (p = 0.039). The relative frequency of CD90⁺SC^ish showed a highly significant inverse correlation with the absolute count of total SC^ish in both, the PB and AP (p = 0.0003 and p = 0.0013 respectively). The relative frequency of CD90⁺SC^ish from the PB also showed a significant (p = 0.0002) inverse relationship with total SC^ish count in the AP. Patients with less than 10% CD90⁺SC^ish in the PB had evidently higher (p = 0.0025) total SC^ishcount in the AP. Conclusion. We speculate that lower CD90⁺SC^ishyield in the AP is not a consequence of an inferior collection efficacy, but most likely a result of several still not fully resolved immature SC cytomorphological/biophysical features. Therefore, following the mobilization by chemotherapy G-CSF, some logical questions appear – whether we should follow the absolute count of total SC^ish, or, whether we should test for relative frequency of CD90⁺SC^ish prior to harvesting. To reach the final conclusions, it is essential to conduct further controlled and larger investigations concerning the correlation of circulating and harvested SCs with patients' hematopoietic recovery.

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