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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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HEMATOPOIESIS

Modulation of in vitro proliferation kinetics and primitive hematopoietic potential of individual human CD34+CD38–/lo cells in G0

Edward F. Srour, Xia Tong, Ki Woong Sung, P. Artur Plett, Susan Rice, Joanne Daggy, Constantin T. Yiannoutsos, Rafat Abonour, and Christie M. Orschell

From the Division of Hematology/Oncology, Department of Medicine, Department of Pediatrics, Herman B Wells Center for Pediatric Research, Department of Microbiology/Immunology, Indiana University Cancer Center, and Department of Biostatistics, Indiana University School of Medicine, Indianapolis, IN.

Whether cytokines can modulate the fate of primitive hematopoietic progenitor cells (HPCs) through successive in vitro cell divisions has not been established. Single human marrow CD34+CD38–/lo cells in the G0 phase of cell cycle were cultured under 7 different cytokine combinations, monitored for proliferation on days 3, 5, and 7, then assayed for long-term culture-initiating cell (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 was independent of cytokine stimulation, suggesting that entry of primitive HPCs into the cell cycle is a 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. Nonobese diabetic/severe combined immunodeficiency (NOD/SCID) transplantation studies revealed analogous results to those obtained with LTC-ICs. These data suggest that although exit from quiescence and commitment to proliferation might be stochastic, kinetics of proliferation, and possibly fate of primitive HPCs, might be modulated by extrinsic factors.


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