Article (Scientific journals)
Functional heterogeneity of human CD34(+) cells isolated in subcompartments of the G0 /G1 phase of the cell cycle.
GOTHOT, André; Pyatt, R.; McMahel, J. et al.
1997In Blood, 90 (11), p. 4384-93
Peer Reviewed verified by ORBi
 

Files


Full Text
Gothot 4384.full.pdf
Publisher postprint (367.42 kB)
Download

All documents in ORBi are protected by a user license.

Send to



Details



Keywords :
Antigens, CD34/analysis; Cell Cycle; Cell Separation; Cells, Cultured; Flow Cytometry; G0 Phase; G1 Phase; Hematopoietic Stem Cells/chemistry/cytology; Humans
Abstract :
[en] Using simultaneous Hoechst 33342 (Hst) and Pyronin Y (PY) staining for determination of DNA and RNA content, respectively, human CD34(+) cells were isolated in subcompartments of the G0 /G1 phase of the cell cycle by flow cytometric cell sorting. In both bone marrow (BM) and mobilized peripheral blood (MPB) CD34(+) cells, primitive long-term hematopoietic culture-initiating cell (LTHC-IC) activity was higher in CD34(+) cells isolated in G0 (G0CD34(+) cells) than in those residing in G1 (G1CD34(+) cells). However, as MPB CD34(+) cells displayed a more homogeneous cell-cycle status within the G0 /G1 phase and a relative absence of cells in late G1 , DNA/RNA fractionation was less effective in segregating LTHC-IC in MPB than in BM. BM CD34(+) cells belonging to four subcompartments of increasing RNA content within the G0 /G1 phase were evaluated in functional assays. The persistence of CD34 expression in suspension culture was inversely correlated with the initial RNA content of test cells. Multipotential progenitors were present in G0 or early G1 subcompartments, while lineage-restricted granulomonocytic progenitors were more abundant in late G1 . In vitro hematopoiesis was maintained for up to 6 weeks with G0CD34(+) cells, whereas production of clonogenic progenitors was more limited in cultures initiated with G1CD34(+) cells. To test the hypothesis that primitive LTHC-ICs would reenter a state of relative quiescence after in vitro division, BM CD34(+) cells proliferating in ex vivo cultures were identified from their quiescent counterparts by a relative loss of membrane intercalating dye PKH2, and were further fractionated with Hst and PY. The same functional hierarchy was documented within the PKH2(dim) population whereby LTHC-IC frequency was higher for CD34(+) cells reselected in G0 after in vitro division than for CD34(+) cells reisolated in G1 or in S/G2 + M. However, the highest LTHC-IC frequency was found in quiescent PKH2(bright) CD34(+) cells. Together, these results support the concept that cells with distinct hematopoietic capabilities follow different pathways during the G0 /G1 phase of the cell cycle both in vivo and during ex vivo culture.
Disciplines :
Hematology
Author, co-author :
GOTHOT, André ;  Centre Hospitalier Universitaire de Liège - CHU > Hématologie biologique et immuno hématologie
Pyatt, R.
McMahel, J.
Rice, S.
Srour, E. F.
Language :
English
Title :
Functional heterogeneity of human CD34(+) cells isolated in subcompartments of the G0 /G1 phase of the cell cycle.
Publication date :
1997
Journal title :
Blood
ISSN :
0006-4971
eISSN :
1528-0020
Publisher :
American Society of Hematology, Washington, United States - District of Columbia
Volume :
90
Issue :
11
Pages :
4384-93
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 08 November 2011

Statistics


Number of views
86 (13 by ULiège)
Number of downloads
289 (2 by ULiège)

Scopus citations®
 
170
Scopus citations®
without self-citations
157
OpenCitations
 
6

Bibliography


Similar publications



Contact ORBi