The low cycling status of mobilized peripheral blood CD34+ cells is not restricted to the more primitive subfraction.
Mobilized peripheral blood progenitor cells (PBPC) have been shown to differ qualitatively from bone marrow (BM) progenitors. The released progenitor cells are predominantly in G0/G1 and show a relatively high percentage of rhodamine dull cells. Within the BM these last two features are characteristic of the more primitive progenitors. Although the mobilized PB cells can give rise to long-term repopulation and thus contain stem cells, the frequency of stem cells is not much higher if long-term initiating cell (LTC-IC) assays are used. To determine whether quiescent stem cells are selectively released or the low-cycle status of PB progenitors is related to the release from the BM microenvironment, the cell cycle status and rhodamine content in the PB and BM during mobilization were studied and compared with steady-state BM. More differentiated and more primitive progenitors were separated based on differentiation markers and cloned in single cell assay. In mobilized PB 54% of the CD34+ cells (n=5) were rhodamine dull compared to 22% in steady-state BM (P=0.014) [n=6]. The percentage of CD34+ cells in the S/G2M phases of the cell cycle was 2.1% in the mobilized PB (n=11), and 18% in steady-state BM (n=11) [P=0.002]. During mobilization the fraction of cells in the S/G2M phase of the cell cycle was 16% in BM (n=7), similar to steady-state BM (P=0.34). The released progenitors represented a selection of BM progenitors, with significantly more primitive progenitors (CD34+/13+/33dim) and less lymphoid precursors (CD34+/19+). Within the more differentiated CD34+113+/33bright, myelomonocytic precursors, both in PB as well as in BM, the percentage S/G2M was relatively higher than in the CD34+/13+/33dim subfraction: in normal BM: median 18% vs 8% (P=0.006) [n=8]; in mobilized PB 3% vs 2% (P=0.03) [n=10]; and in BM during mobilization 24% vs 7% (P=0.01) [n=6]. The cycle status of mobilized PB progenitors was low both in the primitive and more differentiated subfractions. During the mobilization period the BM progenitors are cycling as in steady-state BM. The low-cycle status of the mobilized PB progenitors may be related to the loss of contact with the micro-environment.
Croockewit
A J
AJ
Department of Haematology, University Hospital Nijmegen, The Netherlands.
Raymakers
R A
RA
Smeets
M E
ME
vd Bosch
G
G
Pennings
A H
AH
de Witte
T J
TJ
D018952:Antigens, CD34 / Q000097:blood* / Q000502:physiology*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002458:Cell Fractionation; D002465:Cell Movement / Q000502:physiology; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D016193:G1 Phase / Q000502:physiology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D007202:Indicators and Reagents; D011419:Propidium; D016192:Resting Phase, Cell Cycle / Q000502:physiology; D020112:Rhodamine 123; D012235:Rhodamines / Q000493:pharmacokinetics; D016196:S Phase / Q000502:physiology; D013194:Staining and Labeling / Q000379:methods
D018952:Antigens, CD34; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D012235:Rhodamines; D020112:Rhodamine 123; D011419:Propidium