Efficient generation of CD34+ progenitor-derived dendritic cells from G-CSF-mobilized peripheral mononuclear cells does not require hematopoietic stem cell enrichment

Sophie Paczesny, Yin Ping Li, Na Li, Véronique Latger-Cannard, Luc Marchal, Jing Ping Ou-Yang, Pierre Bordigoni, Jean François Stoltz, Assia Eljaafari

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

As a result of their potent antigen-presentation function, dendritic cells (DC) are important tools for cell therapy programs. In vitro-generated DC from enriched CD34+ hematopoietic stem cells (HSC; enriched CD34 DC) have already proven their efficiency in Phase I/II clinical trials. Here, we investigated whether enrichment of CD34+ HSC before the onset of culture was absolutely required for their differentiation into DC. With this aim, we developed a new two-step culture method. PBMC harvested from G-CSF-mobilized, healthy patients were expanded for 7 days during the first step, with early acting cytokines, such as stem cell factor, fetal liver tyrosine kinase 3 ligand (Flt-3L), and thrombopoietin. During the second step, expanded cells were then induced to differentiate into mature DC in the presence of GM-CSF, Flt-3L, and TNF-α for 8 days, followed by LPS exposure for 2 additional days. Our results showed that the rate of CD34+/CD38+/lineageneg cells increased 19.5 ± 10-fold (mean±SD) during the first step, and the expression of CD14, CD1a, CD86, CD80, and CD83 molecules was up-regulated markedly following the second step. When compared with DC generated from enriched CD34+ cells, which were expanded for 7 days before differentiation, DC derived from nonenriched peripheral blood stem cells showed a similar phenotye but higher yields of production. Accordingly, the allogeneic stimulatory capacity of the two-step-cultured DC was as at least as efficient as that of enriched CD34 DC. In conclusion, we report herein a new two-step culture method that leads to high yields of mature DC without any need of CD34+ HSC enrichment.

Original languageEnglish (US)
Pages (from-to)957-967
Number of pages11
JournalJournal of Leukocyte Biology
Volume81
Issue number4
DOIs
StatePublished - Apr 2007
Externally publishedYes

Fingerprint

Granulocyte Colony-Stimulating Factor
Hematopoietic Stem Cells
Dendritic Cells
fms-Like Tyrosine Kinase 3
Protein-Tyrosine Kinases
Ligands
Thrombopoietin
Phase II Clinical Trials
Clinical Trials, Phase I
Stem Cell Factor
Antigen Presentation
Granulocyte-Macrophage Colony-Stimulating Factor
Cell- and Tissue-Based Therapy
Cultured Cells
Cytokines

Keywords

  • Cell vaccine
  • Ex vivo expansion
  • Immunotherapy
  • Lipopolysaccharide

ASJC Scopus subject areas

  • Cell Biology

Cite this

Efficient generation of CD34+ progenitor-derived dendritic cells from G-CSF-mobilized peripheral mononuclear cells does not require hematopoietic stem cell enrichment. / Paczesny, Sophie; Li, Yin Ping; Li, Na; Latger-Cannard, Véronique; Marchal, Luc; Ou-Yang, Jing Ping; Bordigoni, Pierre; Stoltz, Jean François; Eljaafari, Assia.

In: Journal of Leukocyte Biology, Vol. 81, No. 4, 04.2007, p. 957-967.

Research output: Contribution to journalArticle

Paczesny, Sophie ; Li, Yin Ping ; Li, Na ; Latger-Cannard, Véronique ; Marchal, Luc ; Ou-Yang, Jing Ping ; Bordigoni, Pierre ; Stoltz, Jean François ; Eljaafari, Assia. / Efficient generation of CD34+ progenitor-derived dendritic cells from G-CSF-mobilized peripheral mononuclear cells does not require hematopoietic stem cell enrichment. In: Journal of Leukocyte Biology. 2007 ; Vol. 81, No. 4. pp. 957-967.
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AU - Paczesny, Sophie

AU - Li, Yin Ping

AU - Li, Na

AU - Latger-Cannard, Véronique

AU - Marchal, Luc

AU - Ou-Yang, Jing Ping

AU - Bordigoni, Pierre

AU - Stoltz, Jean François

AU - Eljaafari, Assia

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N2 - As a result of their potent antigen-presentation function, dendritic cells (DC) are important tools for cell therapy programs. In vitro-generated DC from enriched CD34+ hematopoietic stem cells (HSC; enriched CD34 DC) have already proven their efficiency in Phase I/II clinical trials. Here, we investigated whether enrichment of CD34+ HSC before the onset of culture was absolutely required for their differentiation into DC. With this aim, we developed a new two-step culture method. PBMC harvested from G-CSF-mobilized, healthy patients were expanded for 7 days during the first step, with early acting cytokines, such as stem cell factor, fetal liver tyrosine kinase 3 ligand (Flt-3L), and thrombopoietin. During the second step, expanded cells were then induced to differentiate into mature DC in the presence of GM-CSF, Flt-3L, and TNF-α for 8 days, followed by LPS exposure for 2 additional days. Our results showed that the rate of CD34+/CD38+/lineageneg cells increased 19.5 ± 10-fold (mean±SD) during the first step, and the expression of CD14, CD1a, CD86, CD80, and CD83 molecules was up-regulated markedly following the second step. When compared with DC generated from enriched CD34+ cells, which were expanded for 7 days before differentiation, DC derived from nonenriched peripheral blood stem cells showed a similar phenotye but higher yields of production. Accordingly, the allogeneic stimulatory capacity of the two-step-cultured DC was as at least as efficient as that of enriched CD34 DC. In conclusion, we report herein a new two-step culture method that leads to high yields of mature DC without any need of CD34+ HSC enrichment.

AB - As a result of their potent antigen-presentation function, dendritic cells (DC) are important tools for cell therapy programs. In vitro-generated DC from enriched CD34+ hematopoietic stem cells (HSC; enriched CD34 DC) have already proven their efficiency in Phase I/II clinical trials. Here, we investigated whether enrichment of CD34+ HSC before the onset of culture was absolutely required for their differentiation into DC. With this aim, we developed a new two-step culture method. PBMC harvested from G-CSF-mobilized, healthy patients were expanded for 7 days during the first step, with early acting cytokines, such as stem cell factor, fetal liver tyrosine kinase 3 ligand (Flt-3L), and thrombopoietin. During the second step, expanded cells were then induced to differentiate into mature DC in the presence of GM-CSF, Flt-3L, and TNF-α for 8 days, followed by LPS exposure for 2 additional days. Our results showed that the rate of CD34+/CD38+/lineageneg cells increased 19.5 ± 10-fold (mean±SD) during the first step, and the expression of CD14, CD1a, CD86, CD80, and CD83 molecules was up-regulated markedly following the second step. When compared with DC generated from enriched CD34+ cells, which were expanded for 7 days before differentiation, DC derived from nonenriched peripheral blood stem cells showed a similar phenotye but higher yields of production. Accordingly, the allogeneic stimulatory capacity of the two-step-cultured DC was as at least as efficient as that of enriched CD34 DC. In conclusion, we report herein a new two-step culture method that leads to high yields of mature DC without any need of CD34+ HSC enrichment.

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