Degranulation and cytokine expression in human cord blood-derived mast cells cultured in serum-free medium with recombinant human stem cell factor.
Human cord blood-derived mast cells (HCMC) grown in medium with serum and recombinant human stem cell factor (rhSCF) with or without interleukin (IL)-6 are less mature than human skin mast cells (HSMC). We found that c-kit-positive HCMC cultured for 8-10 weeks with rhSCF in serum-free medium became sensitive to basic secretatogues and expressed the serine protease, chymase, which is preferentially expressed in HSMC. The HCMC release beta-hexosaminidase (beta-HEX) within 1 min of stimulation with compound 48/80 or substance P, and release was suppressed by pertussis toxin. Approximately 34% of the HCMC in the serum-free culture stained positively with chymase antibody. Chymase and c-kit levels, and responsiveness to basic secretagogues, increased substantially after an additional 2 weeks in a serum-free environment with rhIL-6 and rhSCF. Moreover, Fc(epsilon)RI-dependent activation of the HCMC resulted in induction of cytokines and cyclooxygenase-2. These results show that HCMC can differentiate into a phenotype morphologically and functionally similar to HSMC if exposed to SCF in serum-free medium.
Moon
Tae Chul
TC
College of Pharmacy, Yeungnam University, Gyongsan 712-749, Korea.
Lee
Eunkyung
E
Baek
Suk-Hwan
SH
Murakami
Makoto
M
Kudo
Ichiro
I
Kim
Nung Soo
NS
Lee
Jong Myung
JM
Min
Hae-Ki
HK
Kambe
Naotomo
N
Chang
Hyeun Wook
HW
D015550:Cell Degranulation / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D053818:Chymases; D016895:Culture Media, Serum-Free / Q000737:chemistry; D051546:Cyclooxygenase 2; D016207:Cytokines / Q000096:biosynthesis* / Q000276:immunology; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology; D007527:Isoenzymes / Q000096:biosynthesis; D008407:Mast Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins; D037342:Pertussis Toxin / Q000494:pharmacology; D011247:Pregnancy; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000096:biosynthesis; D019089:Stem Cell Factor / Q000494:pharmacology*; D013373:Substance P / Q000494:pharmacology; D001619:beta-N-Acetylhexosaminidases / Q000378:metabolism; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D016207:Cytokines; D015850:Interleukin-6; D007527:Isoenzymes; D008565:Membrane Proteins; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D013373:Substance P; D003189:p-Methoxy-N-methylphenethylamine; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D037342:Pertussis Toxin; D001619:beta-N-Acetylhexosaminidases; D012697:Serine Endopeptidases; D053818:Chymases