title: Generation of mast cells from mouse fetus: analysis of differentiation and functionality, and transcriptome profiling using next generation sequencer.
abstract: While gene knockout technology can reveal the roles of proteins in cellular functions, including in mast cells, fetal death due to gene manipulation frequently interrupts experimental analysis. We generated mast cells from mouse fetal liver (FLMC), and compared the fundamental functions of FLMC with those of bone marrow-derived mouse mast cells (BMMC). Under electron microscopy, numerous small and electron-dense granules were observed in FLMC. In FLMC, the expression levels of a subunit of the FcεRI receptor and degranulation by IgE cross-linking were comparable with BMMC. By flow cytometry we observed surface expression of c-Kit prior to that of FcεRI on FLMC, although on BMMC the expression of c-Kit came after FcεRI. The surface expression levels of Sca-1 and c-Kit, a marker of putative mast cell precursors, were slightly different between bone marrow cells and fetal liver cells, suggesting that differentiation stage or cell type are not necessarily equivalent between both lineages. Moreover, this indicates that phenotypically similar mast cells may not have undergone an identical process of differentiation. By comprehensive analysis using the next generation sequencer, the same frequency of gene expression was observed for 98.6% of all transcripts in both cell types. These results indicate that FLMC could represent a new and useful tool for exploring mast cell differentiation, and may help to elucidate the roles of individual proteins in the function of mast cells where gene manipulation can induce embryonic lethality in the mid to late stages of pregnancy.
authors.affiliation: Department of Pharmacology, Faculty of Pharmaceutical Sciences, Tokushima Bunri University, Tokushima, Japan. nobuf@ph.bunri-u.ac.jp
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Fukuishi
authors.forename: Yuusuke
authors.initials: Y
authors.lastname: Igawa
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Kunimi
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Hamano
authors.forename: Masao
authors.initials: M
authors.lastname: Toyota
authors.forename: Hironobu
authors.initials: H
authors.lastname: Takahashi
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Kenmoku
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Yagi
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Matsui
authors.forename: Masaaki
authors.initials: M
authors.lastname: Akagi
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology; D015550:Cell Degranulation; D002454:Cell Differentiation; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005333:Fetus / Q000166:cytology; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D008407:Mast Cells / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019707:Receptors, Chemokine / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013234:Stem Cells / Q000502:physiology*; D059467:Transcriptome
chemical_list: D016207:Cytokines; D012333:RNA, Messenger; D019707:Receptors, Chemokine; D010447:Peptide Hydrolases