Characterization of cord-blood-derived human mast cells cultured in the presence of Steel factor and interleukin-6.
We generated > 10(7) mast cells by culturing 10(7) cord blood mononuclear cells for > 10 weeks in the presence of Steel factor, interleukin-6 and prostaglandin E2. 99% of the cultured cells had tryptase-positive granules, while 18% had chymase-positive granules. Cultured mast cells contained 3.6 micrograms histamine and 3.5 micrograms tryptase per 10(6) cells. Cells sensitized with 1 microgram/ml human IgE released 58.5% histamine and 1.55 ng tumor necrosis factor (TNF)-alpha per 10(6) cells when challenged with 1 microgram/ml antihuman IgE, whereas the control cells spontaneously released 3.7% histamine and 0.18 ng TNF-alpha. Analysis for surface antigens revealed that cultured mast cells expressed the following CD molecules: 9, 13, 14, 29, 33, 38, 43, 44, 45RA, 45RB, 46, 47, 48, 49d, 50, 51, 53, 54, 55, 58, 59, 60, 61 and 117 (c-Kit). Taken together, these cultured cells seem to be functionally mature mast cells.
Saito
H
H
Division of Allergy, National Children's Medical Research Center, Tokyo, Japan.
Ebisawa
M
M
Sakaguchi
N
N
Onda
T
T
Iikura
Y
Y
Yanagida
M
M
Uzumaki
H
H
Nakahata
T
T
D015703:Antigens, CD / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D053818:Chymases; D003594:Cytoplasmic Granules / Q000201:enzymology; D015232:Dinoprostone / Q000494:pharmacology; D005312:Fetal Blood / Q000166:cytology*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology*; D006632:Histamine / Q000494:pharmacology; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology*; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D011994:Recombinant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000032:analysis; D019089:Stem Cell Factor; D053802:Tryptases; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D015703:Antigens, CD; D016298:Hematopoietic Cell Growth Factors; D015850:Interleukin-6; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D014409:Tumor Necrosis Factor-alpha; D007073:Immunoglobulin E; D006632:Histamine; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases; D015232:Dinoprostone