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Title: Establishment and characterization of mouse bone marrow-derived mast cell hybridomas

Abstract

Interleukin (IL)-3-dependent mouse bone marrow-derived mast cells (BMMCs) are an important model for studying the function of mucosal-type mast cells. In the present study, BMMCs were successfully immortalized by cell fusion using a hypoxanthine-aminopterin-thymidine medium-sensitive variant of P815 mouse mastocytoma (P815-6TgR) as a partner cell line. The established mouse mast cell hybridomas (MMCHs) expressed {alpha}, {beta}, and {gamma} subunits of high-affinity immunoglobulin E (IgE) receptor (Fc{epsilon}RI) and possessed cytoplasmic granules devoid of or partially filled with electron-dense material. Four independent MMCH clones continuously proliferated without supplemental exogenous IL-3 and showed a degranulation response on stimulation with IgE+antigen. Furthermore, histamine synthesis and release by degranulation were confirmed in MMCH-D5, a MMCH clone that showed the strongest degranulation response. MMCH-D5 exhibited elevated levels of IL-3, IL-4, IL-13, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor (TNF)-{alpha}, and cyclooxygenase 2, and production of prostaglandin D{sub 2} and leukotriene C{sub 4} in response to IgE-induced stimulation. MMCH clones also expressed Toll-like receptors (TLRs) 1, 2, 4, and 6 and showed elevated levels of TNF-{alpha} expression in response to stimulation with TLR2 and TLR4 ligands. The MMCHs established using this method should be suitable for studies on Fc{epsilon}RI- and TLR-mediated effector functions of mast cells.

Authors:
 [1]
  1. Integrated Department of Sciences of Functional Foods, Graduate School of Agriculture, Shinshu University, Nagano (Japan)
Publication Date:
OSTI Identifier:
22212614
Resource Type:
Journal Article
Journal Name:
Experimental Cell Research
Additional Journal Information:
Journal Volume: 318; Journal Issue: 18; Other Information: Copyright (c) 2012 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0014-4827
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ALBUMINS; BLOOD SERUM; BONE MARROW; CARBOXYPEPTIDASES; CATTLE; DINITROPHENOL; DMSO; FLUORESCEIN; IMMUNOGLOBULINS; LEUKOCYTES; MACROPHAGES; MAST CELLS; MICE; PHOSPHATES; POLYMERASE CHAIN REACTION; RECEPTORS

Citation Formats

Kawahara, Takeshi, E-mail: tkawafb@shinshu-u.ac.jp. Establishment and characterization of mouse bone marrow-derived mast cell hybridomas. United States: N. p., 2012. Web. doi:10.1016/J.YEXCR.2012.07.010.
Kawahara, Takeshi, E-mail: tkawafb@shinshu-u.ac.jp. Establishment and characterization of mouse bone marrow-derived mast cell hybridomas. United States. doi:10.1016/J.YEXCR.2012.07.010.
Kawahara, Takeshi, E-mail: tkawafb@shinshu-u.ac.jp. Thu . "Establishment and characterization of mouse bone marrow-derived mast cell hybridomas". United States. doi:10.1016/J.YEXCR.2012.07.010.
@article{osti_22212614,
title = {Establishment and characterization of mouse bone marrow-derived mast cell hybridomas},
author = {Kawahara, Takeshi, E-mail: tkawafb@shinshu-u.ac.jp},
abstractNote = {Interleukin (IL)-3-dependent mouse bone marrow-derived mast cells (BMMCs) are an important model for studying the function of mucosal-type mast cells. In the present study, BMMCs were successfully immortalized by cell fusion using a hypoxanthine-aminopterin-thymidine medium-sensitive variant of P815 mouse mastocytoma (P815-6TgR) as a partner cell line. The established mouse mast cell hybridomas (MMCHs) expressed {alpha}, {beta}, and {gamma} subunits of high-affinity immunoglobulin E (IgE) receptor (Fc{epsilon}RI) and possessed cytoplasmic granules devoid of or partially filled with electron-dense material. Four independent MMCH clones continuously proliferated without supplemental exogenous IL-3 and showed a degranulation response on stimulation with IgE+antigen. Furthermore, histamine synthesis and release by degranulation were confirmed in MMCH-D5, a MMCH clone that showed the strongest degranulation response. MMCH-D5 exhibited elevated levels of IL-3, IL-4, IL-13, granulocyte-macrophage colony-stimulating factor, tumor necrosis factor (TNF)-{alpha}, and cyclooxygenase 2, and production of prostaglandin D{sub 2} and leukotriene C{sub 4} in response to IgE-induced stimulation. MMCH clones also expressed Toll-like receptors (TLRs) 1, 2, 4, and 6 and showed elevated levels of TNF-{alpha} expression in response to stimulation with TLR2 and TLR4 ligands. The MMCHs established using this method should be suitable for studies on Fc{epsilon}RI- and TLR-mediated effector functions of mast cells.},
doi = {10.1016/J.YEXCR.2012.07.010},
journal = {Experimental Cell Research},
issn = {0014-4827},
number = 18,
volume = 318,
place = {United States},
year = {2012},
month = {11}
}