Generation and functional characterization of human dendritic cells derived from CD34+ cells mobilized into peripheral blood: Comparison with bone marrow CD34+ cells

Marina Ratta, Damiano Rondelli, Alessandra Fortuna, Antonio Curti, Miriam Fogli, Francesco Fagnoni, Giovanni Martinelli, Carolina Terragna, Sante Tura, Roberto M. Lemoli

Research output: Contribution to journalArticlepeer-review


Dendritic cells (DCs) are the most powerful professional antigen- presenting cells (APC), specializing in capturing antigens and stimulating T- cell-dependent immunity. In this study we report the generation and characterization of functional DCs derived from both steadystate bone marrow IBM) and circulating haemopoietic CD34+ cells from 14 individuals undergoing granulocyte colony-stimulating factor (G-CSF) treatment for peripheral blood stem cells (PBSC) mobilization and transplantation. Clonogenic assays in methylcellulose showed an increased frequency and proliferation of colony- forming unit-dendritic cells (CFU-DC) in circulating CD34+ cells, compared to that of BM CD34+ precursors in response to GM-CSF and TNF-α with or without SCF and FLT-3L. Moreover, peripheral blood (PB) CD34+ cells generated a significantly higher number of fully functional DCs, as determined by conventional mixed lymphocyte reactions (MLR), than their BM counter-parts upon different culture conditions. DCs derived from mobilized stem cells were also capable of processing and presenting soluble antigens to autologous T cells for both primary and secondary immune response. Replacement of the early-acting growth factors SCF and FLT-3L with IL-4 at day 7 of culture of PB CD34+ cells enhanced both the percentage of total CD1a+ cells and CD1a+CD14- cells and the yield of DCs after 14d of incubation. In addition, the alloreactivity of IL-4-stimulated DCs was significantly higher than those generated in the absence of IL-4. Furthermore, autologous serum collected during G-CSF treatment was more efficient than fetal calf serum (FCS) or two different serum-free media for large-scale production of DGs. Thus, our comparative studies indicate that G- CSF mobilizes CD34+ DC precursors into PB and circulating CD34+ cells represent the optimal source for the massive generation of DCs. The sequential use of early-acting and intermediatelate-acting colony-stimulating factors (CSFs) as well as the use of autologous serum greatly enhanced the growth of DCs. These data may provide new insights for manipulating immunocompetent cells for cancer therapy.

Original languageEnglish
Pages (from-to)756-765
Number of pages10
JournalBritish Journal of Haematology
Issue number4
Publication statusPublished - 1998


  • CD34 cells
  • Dendritic cells
  • Immunotherapy
  • Peripheral blood haemopoietic progenitors

ASJC Scopus subject areas

  • Hematology


Dive into the research topics of 'Generation and functional characterization of human dendritic cells derived from CD34+ cells mobilized into peripheral blood: Comparison with bone marrow CD34+ cells'. Together they form a unique fingerprint.

Cite this