Generation of a large number of connective tissue type mast cells by culture of murine fetal skin cells.
We describe a novel culture system for generating large numbers of murine skin-associated mast cells and distinguish their characteristics from bone marrow-derived cultured mast cells. Culture of day 16 fetal skin single cell suspensions in the presence of interleukin-3 and stem cell factor allowed expansion and maturation of mast cells in the presence of stromal cells. The average yield of mast cells after 2 wk was 7.3 million cells per fetus at a purity of 96%. These fetal skin-derived cultured mast cells increased their histamine content in a time-dependent manner to 3.6 pg per cell after 2 wk and 6.7 pg per cell after 4 wk. Phenotypic analyses revealed much greater expression of CD49b and CD81 and lesser expression of CD77 and CD102 on fetal skin-derived cultured mast cells as compared with bone marrow-derived cultured mast cells. These findings suggest a close similarity between fetal skin-derived cultured mast cells and freshly isolated cutaneous mast cells. Connective tissue mast cell characteristics of fetal skin-derived cultured mast cells were evidenced by: (1) their greater histamine content than bone marrow-derived cultured mast cells; (2) the presence of heparin; and (3) their degranulation in response to compound 48/80 and substance P. Importantly, fetal skin-derived cultured mast cells secreted greater amounts of interleukin-13 but much less MIP-1beta and interleukin-6 than bone marrow-derived cultured mast cells in response to ionomycin. Thus fetal skin-derived cultured mast cells have many characteristics distinct from bone marrow-derived cultured mast cells and can be used as a model of cutaneous mast cells to discern their functions.
Yamada
Nobuo
N
Dermatology Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
Matsushima
Hironori
H
Tagaya
Yutaka
Y
Shimada
Shinji
S
Katz
Stephen I
SI
D000367:Age Factors; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D015550:Cell Degranulation / Q000187:drug effects; D002454:Cell Differentiation; D002455:Cell Division; D002478:Cells, Cultured; D054407:Chemokine CCL4; D005333:Fetus / Q000166:cytology; D006493:Heparin / Q000378:metabolism; D006632:Histamine / Q000378:metabolism; D018793:Interleukin-13 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D019402:Macrophage Inflammatory Proteins / Q000378:metabolism; D008407:Mast Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018945:Monocyte Chemoattractant Proteins / Q000235:genetics; D010641:Phenotype; D012333:RNA, Messenger / Q000032:analysis; D017455:Receptors, IgE / Q000378:metabolism; D012867:Skin / Q000166:cytology*; D017154:Stromal Cells / Q000166:cytology; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
D054407:Chemokine CCL4; D018793:Interleukin-13; D015850:Interleukin-6; D019402:Macrophage Inflammatory Proteins; D018945:Monocyte Chemoattractant Proteins; D012333:RNA, Messenger; D017455:Receptors, IgE; D003189:p-Methoxy-N-methylphenethylamine; D006632:Histamine; D006493:Heparin