title: Fibroblasts feeder niche and Flt3 Ligand as a novel inducer of plasmacytoid dendritic cells development in vitro.
abstract: Plasmacytoid dendritic cell (pDC), plays central role in antiviral immunity. The aim of this study was to assess the effect of Flt3 ligand (FL) alone or with L929 fibroblast feeder or L929 conditioned media on differentiation of mouse bone marrow (BM) cells into pDC in vitro. Murine BM cells were cultured with FL or with L929 or conditioned media for 9days. The differentiated cells were analyzed using flow cytometry for PDCA-1, B220 and CXCR4. The relative expression of Stat3, CXCR4, CXCR7, IFN-β, TGF-β and Runx2 in differentiated cells determined by real time PCR. The development of pDC showed up to 19% increase after co-culture of BM cells with fibroblast feeder. Upregulation of Stat3, Runx2 and CXCR4 due to the presence of fibroblast feeder with FL in culture results in improved pDC development. Furthermore, 30% L929 supernatant along with Flt3 ligand was able to derive pDC up to 8.9% in comparison with FL alone, which was 6.6% in vitro. Thus, for the first time we introduced L929 fibroblast feeder as a niche producer of M-CSF and probably other growth factors and chemokines, which promotes the development of pDC in vitro along with FL, similar to in vivo niche.
authors.affiliation: Department of Immunology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.
authors.forename: Mohammad Taher
authors.initials: MT
authors.lastname: Tahoori
authors.affiliation: Department of Immunology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran. Electronic address: pourfa@modares.ac.ir.
authors.forename: Ali Akbar
authors.initials: AA
authors.lastname: Pourfathollah
authors.affiliation: Department of Stem Cell Biology, Stem Cell Technology Research Center, Tehran Iran; Department of Hematology, Faculty of Medical Science, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Chemical Engineering, Biotechnology Division, Tarbiat Modares University, Tehran, Iran.
authors.forename: Ebrahim
authors.initials: E
authors.lastname: Vasheghani-Farahani
authors.affiliation: Department of Microbiology, Immunology and Genetics, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.
authors.forename: Adel
authors.initials: A
authors.lastname: Mohammadzadeh
authors.affiliation: Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
authors.forename: Afshin
authors.initials: A
authors.lastname: Amari
authors.affiliation: Department of Immunology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Seyed Mahmoud
authors.initials: SM
authors.lastname: Hashemi
authors.affiliation: Department of Stem Cell Biology, Stem Cell Technology Research Center, Tehran Iran.
authors.forename: Majid
authors.initials: M
authors.lastname: Mossahebi-Mohammadi
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D018920:Coculture Techniques; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D003713:Dendritic Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D008565:Membrane Proteins / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D019718:Receptors, CXCR4 / Q000235:genetics; D050796:STAT3 Transcription Factor / Q000235:genetics
chemical_list: C514936:CXCR4 protein, mouse; D050659:Core Binding Factor Alpha 1 Subunit; D008565:Membrane Proteins; D019718:Receptors, CXCR4; C493716:Runx2 protein, mouse; D050796:STAT3 Transcription Factor; C494087:Stat3 protein, mouse; C084881:flt3 ligand protein; D016178:Granulocyte-Macrophage Colony-Stimulating Factor
keywords: Flt3 ligand; L929 fibroblast feeder; Plasmacytoid dendritic cell