Molecular and stimulus‐response profiles illustrate heterogeneity between peripheral and cord blood‐derived human mast cells

[摘要] DifferentprotocolsexistforinvitrodevelopmentofHuMCsfromhematopoieticstemcells,whichresultsindistinctmastcellsregardingmolecularmarkersandactivationpatterns.Here,weintroduceaSRprofileusingimmunological,neurogenic,andpharmacologicalstimulitocharacterizecellularfunctionality.MastcellswereobtainedfromthreecultureprotocolsusingtwotypesofPBdMCs(CD34+PBdMCorCD133+PBdMC)andonetypeofCBdMC(CD133+CBdMC).WeanalyzedrestingcellsforspecificmastcellmarkersatproteinandmRNAlevels,therebycreatingamolecularprofile.TocharacterizetheSRprofile,westimulatedcellswithanti‐IgE,C3a,C5a,SubstanceP,orCompound48/80andmeasuredthereleaseofhistamineandcytokines(IL‐10,IL‐13,GM‐CSF,TNF‐α).MolecularprofilingrevealedthatCD133+CBdMCexpressedlesschymase,FcɛRIα,andCD203cbutmoreCD117comparedwithCD34+andCD133+PBdMC.TheSRprofileforhistaminereleaseillustratedafunctionalheterogeneitybetweenPBdMCandCBdMC.PBdMCreleased>10%histamineuponstimulationwithanti‐IgE,C3a,SubstanceP,andCompound48/80,whereasCBdMConlyreactedtoC3a.Cytokinesecretionwasonlydetectedafteranti‐IgEstimulation.Here,theSRprofileidentifiedtheCD133+PBdMCasthemostactivecellsregardingsecretionofIL‐10,IL‐13,GM‐CSF,andTNF‐α.Cellsfromallthreecultureprotocols,however,producedIL‐10spontaneouslyatcomparablelevels.WerecommendvalidatingmastcellculturesbymeansofmolecularandSRprofilestocharacterizethemastcellsandenhanceconsensusamongstudies...

[学科分类] 生理学

[关键词] IgE;complement;histamine;cytokine;stem cells