title: A novel method to generate and culture human mast cells: Peripheral CD34+ stem cell-derived mast cells (PSCMCs).
abstract: The identification and characterization of human mast cell (MC) functions are hindered by the shortage of MC populations suitable for investigation. Here, we present a novel technique for generating large numbers of well differentiated and functional human MCs from peripheral stem cells (=peripheral stem cell-derived MCs, PSCMCs). Innovative and key features of this technique include 1) the use of stem cell concentrates, which are routinely discarded by blood banks, as the source of CD34+ stem cells, 2) cell culture in serum-free medium and 3) the addition of LDL as well as selected cytokines. In contrast to established and published protocols that use CD34+ or CD133+ progenitor cells from full blood, we used a pre-enriched cell population obtained from stem cell concentrates, which yielded up to 10(8) differentiated human MCs per batch after only three weeks of culture starting with 10(6) total CD34+ cells. The total purity on MCs (CD117+, FcεR1+) generated by this method varied between 55 and 90%, of which 4-20% were mature MCs that contain tryptase and chymase and show expression of FcεRI and CD117 in immunohistochemistry. PSCMCs showed robust histamine release in response to stimulation with anti-FcεR1 or IgE/anti-IgE, and increased proliferation and differentiation in response to IL-1β or IFN-γ. Taken together, this new protocol of the generation of large numbers of human MCs provides for an innovative and suitable option to investigate the biology of human MCs.
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany. Electronic address: oliver.schmetzer@gmx.net.
authors.forename: Oliver
authors.initials: O
authors.lastname: Schmetzer
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany.
authors.forename: Patricia
authors.initials: P
authors.lastname: Valentin
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany.
authors.forename: Anna
authors.initials: A
authors.lastname: Smorodchenko
authors.affiliation: Department Medical and Biological Sciences, Section of Surgical Pathology, University of Udine, Italy.
authors.forename: Rossana
authors.initials: R
authors.lastname: Domenis
authors.affiliation: Department Medical and Biological Sciences, Section of Surgical Pathology, University of Udine, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Gri
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany.
authors.forename: Frank
authors.initials: F
authors.lastname: Siebenhaar
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Metz
authors.affiliation: Dept. of Dermatology and Allergy, Charité - Universitätsmedizin Berlin, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Maurer
mesh_terms: D000071916:AC133 Antigen; D000906:Antibodies / Q000494:pharmacology; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D018952:Antigens, CD34 / Q000235:genetics* / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D010455:Peptides / Q000235:genetics / Q000276:immunology; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000276:immunology; D017455:Receptors, IgE / Q000235:genetics / Q000276:immunology
chemical_list: D000071916:AC133 Antigen; D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D015703:Antigens, CD; D018952:Antigens, CD34; D015415:Biomarkers; D016895:Culture Media, Serum-Free; D006023:Glycoproteins; C508596:IL3 protein, human; D007377:Interleukin-3; D008077:Lipoproteins, LDL; C000606023:PROM1 protein, human; D010455:Peptides; D017455:Receptors, IgE; C026408:anti-IgE antibodies; D007073:Immunoglobulin E; D019009:Proto-Oncogene Proteins c-kit
keywords: Culture; Differentiation; IgE; Immunoglobulin E; In vitro; Mast cell