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Development of dendritic cells in vitro from murine fetal
liver–derived lineage phenotype-negative c-kit
+hematopoietic progenitor cells
by Yanyun Zhang, Yi Zhang, Yong Wang, Masafumi Ogata, Shin-ichi Hashimoto,
Nobuyuki Onai, and Kouji Matsushima
Blood
Volume 95(1):138-146
January 1, 2000
©2000 by American Society of Hematology
Schematic representation of culture conditions of purified murine FL-derived Lin− c-kit + HPCs.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
The culture of murine 13 dpc FL-derived Lin− c-kit + HPCs.(A) Using a phase contrast
microscope, morphologic analyses were performed on cultured murine 13 dpc FL-derived Lin−
c-kit + HPCs stimulated with GM-CSF + SCF + Flt3L + mTNFα at day 7, 14, and 21.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Generation of DCs from 13dpc FL-derived Lin− c-kit + HPCs.13 dpc FL-derived Lin− c-kit + HPCs
were cocultured with stromal cell lines PA6 or OP9 cells in the presence of GM-CSF + SCF +
Flt3L for 12 to 14 days, and restimulated by GM-CSF + mTNFα for an addit...
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Immunophenotypic and morphologic analyses of DC-like cells.13dpc FL-derived Lin− c-kit +
HPCs were first supplemented with PA6 cells + GM-CSF + SCF + Flt3L for 12 to 14 days and then
stimulated with GM-SCF + mTNFα for an additional 3 to 5 days.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
The capacity of the cultured cells to enhance allogeneic MLR.Allogeneic MLR was performed
using purified T cells (3 × 105 cells/well in 96-round-well plate) as responder cells.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
The role of stromal cells in the induction of DCs from FL-derived Lin− c-kit + HPCs.13 dpc FLderived HPCs were cultured in the presence of cytokines, GM-CSF + SCF + Flt3L, with or without
stromal cell line OP9 or PA6 cells in a transwell culture system.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Immunofluorescence analysis on DC precursors generated from murine FL-derived Lin− c-kit
+HPCs.Immunofluorescence analysis was performed on the nonadherent cells from FL-derived
HPCs cocultured with PA6 cells + GM-CSF + SCF + Flt3L for 12 to 14 days.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Endocytic activity of FL HPC-derived DC precursors and mature DCs.The capacity of FITC-DX
uptake was analyzed by a cell sorter as described in “Materials and Methods.” The FITC
intensity of cells as a control or the test of FITC-DX uptake was indicated.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Generation of NK1.1+ cells from FL-derived Lin− c-kit + HPCs.13 dpc FL-derived Lin− c-kit +
HPCs were cocultured with PA6 cells in the presence or absence of SCF + Flt3L with or without
GM-CSF in a transwell culture system.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Immunophenotypic analysis on NK1.1+ cells generated from FL-derived Lin− c-kit + HPCs.Lin−
c-kit + HPCs from 13 dpc FL were cultured in the presence of PA6 cells + SCF + Flt3L for 14
days.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology
Capacity to generate DC precursors from NK1.1+ cells.13 dpc FL-derived HPCs were co-cultured
with PA6 cells + SCF + Flt3L in transwell plates.
Yanyun Zhang et al. Blood 2000;95:138-146
©2000 by American Society of Hematology