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Blood, 15 April 2005, Vol. 105, No. 8, pp. 3109-3116.
Prepublished online as a Blood First Edition Paper on December 21, 2004; DOI 10.1182/blood-2004-05-1773.
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Submitted May 11, 2004
Accepted December 13, 2004
Modulation of in vitro proliferation kinetics and primitive hematopoietic potential of individual human CD34+CD38-/lo cells ING0
Edward F Srour*, Xia Tong, Ki Woong Sung, P A Plett, Susan Rice, Joanne Daggy, Constantin T Yiannoutsos, Rafat Abonour, and Christie M Orschell
Division of Hematology/Oncology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Microbiology/Immunology, Indiana University School of Medicine, Indianapolis, IN, USA
Division of Hematology/Oncology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA
Department of Biostatistics, Indiana University School of Medicine, Indianapolis, IN, USA
Indiana University Cancer Center, Indianapolis, IN, USA; Department of Biostatistics, Indiana University School of Medicine, Indianapolis, IN, USA
Division of Hematology/Oncology, Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA; Indiana University Cancer Center, Indianapolis, IN, USA
* Corresponding author; email: esrour{at}iupui.edu.
Whether cytokines can modulate the fate of primitive hematopoietic progenitor cells (HPC) through successive in vitro cell divisions has not been established. Single human marrow CD34+CD38-/lo cells in G0 phase of cell cycle were cultured under 7 different cytokine combinations, monitored for proliferation on days 3, 5 and 7, then assayed for LTC-IC function on day 7. LTC-IC function was then retrospectively correlated with prior number of in vitro cell divisions to determine whether maintenance of LTC-IC function after in vitro cell division is dependent on cytokine exposure. In the presence of proliferation progression signals, initial cell division on days 3 and 5 was independent of cytokine stimulation, suggesting that entry of primitive HPC into cell cycle is an intrinsic stochastic property. However, kinetics of proliferation beyond day 3 and maintenance of LTC-IC function were sensitive to cytokine stimulation, such that LTC-IC underwent an initial long cell cycle, followed by more synchronized shorter cycles varying in length depending on the cytokine combination. In vitro observations were supported by NOD/SCID transplantation studiesNOD/SCID transplantation studies revealed analogus results to those obtained with LTC-IC. These data suggest that while exit from quiescence and commitment to proliferation might be deterministicstochastic, kinetics of proliferation, and possibly and fate of primitive HPC,, including repopulating cells, might be controlled modulated by extrinsic factors.
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