Development and functional analysis of eosinophils from murine embryonic stem cells.
We have established a culture system for the development of eosinophils from murine embryonic stem (ES) cells. After transferring ES cells from embryonic fibroblast cells onto macrophage colony-stimulating factor-deficient stromal cells, OP9, ES cells were cultured in the presence of interleukin (IL)-5 with either IL-3 or granulocyte-macrophage colony stimulating factor (GM-CSF) for 20 d to obtain approximately 50% eosinophils. Electron microscopy confirmed the presence of crystallized major basic protein (MBP) in the granules of some of these cells. Neither IL-5, IL-3, GM-CSF nor eotaxin alone could induce eosinophils as efficiently as the conditions described above. Eotaxin induced eosinophil development in combination with either IL-3 or IL-5. Levels of GATA-1, Friend of GATA (FOG)-1, PU.1, CCAAT/enhancer binding protein (C/EBP)alpha, C/EBPbeta, IL-3 receptor alpha (IL-3Ralpha), GM-CSF receptor alpha (GM-CSFRalpha), and MBP mRNAs were increased in ES cells 10 d after transfer onto OP9 cells. In contrast, C/EBPepsilon, IL-5Ralpha, and eosinophil peroxidase mRNAs were induced in response to IL-3 and IL-5 after transfer onto OP9 cells. Eosinophils that developed in this system expressed Gr-1, F4/80, B220, CCR3, IL-3Ralpha, IL-5Ralpha, and DX5. Finally, eosinophils developed from ES cells produced reactive oxygen species in response to Leishmania as do peripheral blood eosinophils.
Hamaguchi-Tsuru
Emi
E
Department of Molecular and Cellular Biology, Kochi Medical School, Nankoku City, Kochi, Japan.
Nobumoto
Atsuya
A
Hirose
Noriyuki
N
Kataoka
Sayo
S
Fujikawa-Adachi
Kiyomi
K
Furuya
Masato
M
Tominaga
Akira
A
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003460:Crystallization; D016207:Cytokines / Q000494:pharmacology; D004622:Embryo, Mammalian / Q000166:cytology*; D047091:Eosinophil Granule Proteins; D047209:Eosinophil Peroxidase; D004804:Eosinophils / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron; D010544:Peroxidases / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012260:Ribonucleases / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D015854:Up-Regulation
D000954:Antigens, Surface; D015415:Biomarkers; D001798:Blood Proteins; D016207:Cytokines; D047091:Eosinophil Granule Proteins; D017382:Reactive Oxygen Species; D047209:Eosinophil Peroxidase; D010544:Peroxidases; D012260:Ribonucleases