Cultivation and characterization of canine skin-derived mast cells.
It is essential to develop a technique to culture purified skin-derived mast cells (SMCs) to facilitate immunological research on allergic diseases in dogs. This study was performed to develop an efficient culture system for canine SMCs and to characterize the cells in comparison to canine bone marrow-derived mast cells (BMMCs). Enzymatically digested skin biopsy samples were cultivated in serum-free AIM-V medium supplemented with recombinant canine stem cell factor. Three to five weeks after the initiation of culture, mast cells were collected by a magnetic activated cell separation system using anti-c-Kit antibody. The collected cells were composed of a uniform population showing morphological characteristics of mast cells with a round or oval nucleus and abundant toluidine blue-positive metachromatic granules in the cytoplasm. The results of flow cytometric analysis for the presence of cell membrane c-Kit and Fc epsilon receptor I (FcepsilonRI) indicated that approximately 90% of the cells were mast cells. The cytoplasmic granules were positive for both tryptase and chymase. Apparent dose-dependent degranulation was induced by antibody-mediated cross-linking of immunoglobulin E (IgE) bound to the cells. These cytological and immunological characteristics observed in SMCs were mostly similar to those observed in BMMCs; however, IgE-mediated degranulation was significantly lower in SMCs than BMMCs. The culture system for canine SMCs developed in this study would be useful in understanding the pathophysiology and developing anti-allergic therapeutics in canine allergic dermatitis.
Kawarai
Shinpei
S
First Department of Veterinary Microbiology, School of Veterinary Medicine, Azabu University, 1-7-71 Fuchinobe, Sagamihara, Kanagawa 229-8501, Japan.
Masuda
Kenichi
K
Ohmori
Keitaro
K
Matsuura
Shinobu
S
Yasuda
Nobutaka
N
Nagata
Masahiko
M
Sakaguchi
Masahiro
M
Tsujimoto
Hajime
H
D000818:Animals; D001399:Azure Stains / Q000737:chemistry; D001560:Benzidines / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary*; D053818:Chymases / Q000032:analysis; D017449:Dermatitis, Allergic Contact / Q000276:immunology / Q000662:veterinary*; D004285:Dogs; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D007150:Immunohistochemistry / Q000662:veterinary; D008297:Male; D008407:Mast Cells / Q000166:cytology / Q000201:enzymology / Q000276:immunology* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission / Q000662:veterinary; D019009:Proto-Oncogene Proteins c-kit / Q000032:analysis; D017455:Receptors, IgE / Q000032:analysis; D012867:Skin / Q000166:cytology / Q000276:immunology* / Q000648:ultrastructure; D053802:Tryptases / Q000032:analysis; D001619:beta-N-Acetylhexosaminidases / Q000032:analysis
D001399:Azure Stains; D001560:Benzidines; D017455:Receptors, IgE; C009501:2-tolidine; D019009:Proto-Oncogene Proteins c-kit; D001619:beta-N-Acetylhexosaminidases; D053818:Chymases; D053802:Tryptases