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(Journal of Leukocyte Biology. 2002;72:321-329.)
© 2002 by Society for Leukocyte Biology

Expansion of dendritic cell precursors from human CD34+ progenitor cells isolated from healthy donor blood; growth factor combination determines proliferation rate and functional outcome

Hetty J. Bontkes*, Tanja D. de Gruijl{dagger}, Gert Jan Schuurhuis{ddagger}, Rik J. Scheper*, Chris J. L. M. Meijer* and Erik Hooijberg*

Departments of
* Pathology,
{dagger} Medical Oncology, and
{ddagger} Hematology, Vrije Universiteit Medical Center, Amsterdam, The Netherlands

Correspondence: H. J. Bontkes, Ph.D., Dept. of Pathology, Vrije Universiteit Medical Center, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands. E-mail: hj.bontkes{at}vumc.nl

CD34+ haematopoietic progenitor cells, which circulate at extremely low frequencies in peripheral blood, are used to generate dendritic cells (DC) in vitro. Here, we describe a method to grow large numbers of DC precursors from these low frequent cells. Different combinations of early acting haematopoietic growth factors supported expansion of CD34+ cells. CD1a+ DC derived from precursors, expanded in fms-like tyrosine kinase-3 ligand (Flt3-L), stem-cell factor (SCF), interleukin (IL)-3, and IL-6, were less potent antigen-presenting cells (APC) compared to CD1a+ DC derived from precursors expanded in Flt3-L, trombopoietine (TPO), and SCF. Furthermore, the latter produced high levels of IL-12 and low levels of IL-10, a cytokine profile favorable for the priming cytotoxic T cells. In contrast, a mean increase of total cell number of 453-fold was obtained with Flt3-L, SCF, IL-3, and IL-6, and this increase was only 38-fold with Flt3-L, TPO, and SCF. Sequential cultures of both cocktails resulted in high numbers of potent APC, which can be useful DC-based cancer vaccines.

Key Words: tumor immunity • vaccination • cytokines




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