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Blood, Vol. 95 No. 1 (January 1), 2000:
pp. 138-146
Development of dendritic cells in vitro from murine fetal
liver-derived lineage phenotype-negative c-kit+
hematopoietic progenitor cells
Yanyun Zhang,
Yi Zhang,
Yong Wang,
Masafumi Ogata,
Shin-ichi Hashimoto,
Nobuyuki Onai, and
Kouji Matsushima
From the Department of Molecular Preventive Medicine and CREST,
School of Medicine, The University of Tokyo, Japan.
We describe here that lineage phenotype- negative
(Lin) c-kit+ hematopoietic
progenitor cells (HPCs) from day 13 postcoitus (dpc) murine fetal liver
(FL) can generate dendritic cell (DC) precursors when cultured in vitro
in the presence of PA6 stromal cells plus granulocyte/macrophage
colony-stimulating factor (GM-CSF) + stem cell factor (SCF) + Flt3
ligand (Flt3L) for 12 to 14 days, and develop into mature DCs when
stimulated with GM-CSF plus mouse tumor necrosis factor  (mTNF)
for an additional 3 to 5 days. A transwell culture system showed
that the generation of DC precursors depended on the support of PA6
cell-secreted soluble factor(s). The mature DCs derived from 13 dpc FL
Linc-kit+ HPCs showed
characteristic morphology and function of DCs and expressed high levels
of Ia, CD86, and CD40 molecules, low levels of DEC205, E-cadherin, and
F4/80 molecules, but barely detectable CD11c antigen. Once FL-derived
HPCs were cultured without GM-CSF, NK1.1+ cells developed
in the presence of PA6 cells + SCF + Flt3L. These NK1.1+ cells could develop into DC precursors at an
earlier stage of differentiation by reculturing with PA6 cells + SCF + Flt3L + GM-CSF, but they would be irreversibly
committed to NK cell precursors without GM-CSF after 3 days,
suggesting that GM-CSF plays a critical role in controlling the
transition of DC and NK cell precursors from 13 dpc FL-derived
Linc-kit+ HPCs. This study
represents the first success in generating mature DCs in vitro from
murine FL HPCs. (Blood. 2000;95:138-146)
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