An improved procedure for the development of human mast cells from dispersed fetal liver cells in serum-free culture medium

Kambe, Naotomo; Kambe, Michiyo; Chang, Hyeun‐Wook; Matsui, Atsushi; Min, Hae‐Ki; Hussein, Mousa I.; Oskerizian, Carole A; Kochan, Jarko; Irani, Anne-Marie; Schwartz, Lawrence B.

doi:10.1016/s0022-1759(00)00174-5

Cited by 32 publications

(36 citation statements)

References 18 publications

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“…23 For chymase detection, B7-biotin-labeled cells were incubated with peroxidase-conjugated streptavidin and visualized with 3-amino-9-ethylcarbazole in 0.01% H 2 O 2 to stain positive cells reddish brown. For tryptase detection, alkaline phosphatase-conjugated goat antimouse IgG (Sigma) was added to G3-labeled cells and visualized with naphthol AS-BI/new fuchsin to stain positive cells red.…”

Section: Immunochemistrymentioning

confidence: 99%

“…Serum-free AIM-V media permitted an earlier, more selective and greater expansion of fetal liver-derived mast cells than serum-containing media in the presence of rhuSCF. 23 However, most tissue-derived human mast cells, though probably long-lived, are thought to be terminally differentiated with limited, if any, proliferative capacity. The growth of skin-derived mast cells in the presence of rhuSCF (100 ng/mL) was assessed with this serum-free media compared to serum-containing RPMI 1640 ( Figure 1).…”

Section: Proliferation Of Human Skin-derived Mast Cells In a Serum-frmentioning

confidence: 99%

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Human skin–derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes

Kambe

Kochan

et al. 2001

Blood

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IntroductionMultipotential progenitor cells, capable of becoming mast cells, leave the bone marrow and enter the circulation, but they complete their differentiation into mature mast cells only after arriving in peripheral tissues such as lung, bowel, skin, and nasal and conjunctival mucosa. Mature human mast cells can be distinguished from other cell types by expression of high levels of surface Fc⑀RI and Kit, and of granule tryptase and heparin. Two types of human mast cells have been identified based on their composition of neutral proteases. 1-6 MC TC cells contain tryptase, chymase, mast cell carboxypeptidase, and cathepsin G in their secretory granules and are the predominant type of mast cells in normal skin and in intestinal submucosa. MC T cells also contain tryptase in their granules, but lack these other proteases, and are the predominant type of mast cell in normal alveolar wall and in small intestinal mucosa.In a number of human diseases such as asthma, 7,8 allergic rhinitis, 9,10 rheumatoid arthritis, [11][12][13] and vernal keratoconjunctivitis, 14 significant increases in mast cell density at the local affected sites have been described and mast cells were estimated to play a central role in the pathophysiology of the associated inflammation. The principal mechanisms involved in mast cell hyperplasia were thought to be either the recruitment of progenitor cells and their differentiation to mast cells or the migration of mast cells from other regions of the tissue. Proliferation has been considered less likely in part because cells were considered to be terminally differentiated and also because mitotic mast cells are rarely appreciated at these sites.The proliferative potential of most myelomonocytic multipotential hematopoietic cells diminishes as they differentiate. However, mature murine mast cells have been reported to proliferate both in vivo and in vitro. About 6% of mouse peritoneal mast cells proliferate after being injected into skin of W/W v mast cell-deficient mice. 15,16 Purified peritoneal mast cells 17,18 or dispersed skin mast cells, 19 when plated in methylcellulose containing pokeweed mitogen-stimulated spleen cell-conditioned medium, produced mast cell colonies. However, in humans, isolated tissue-derived mast cells have shown little capacity for proliferation. For example, dispersed newborn foreskin or adult skin mast cells placed into culture with lymphocyte-conditioned media and various cytokines showed bromodeoxyuridine incorporation in up to 15% mast cells. 20 The current study shows that a substantial portion of mast cells dispersed from adult human skin can proliferate under serum-free culture conditions with recombinant human stem cell factor (rhuSCF), while retaining expression of chymase and tryptase in secretory granules, Kit, and Fc⑀RI on their surface and the ability to degranulate in response to IgG anti-Fc⑀RI␣, compound 48/80 and substance P. Materials and methods Antibodies and reagentsBiotin-conjugated mouse IgG monoclonal antibody (mAb) antichymase, B7-biotin, ...

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Section: Immunochemistrymentioning

confidence: 99%

Section: Proliferation Of Human Skin-derived Mast Cells In a Serum-frmentioning

confidence: 99%

Human skin–derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes

Kambe

Kochan

et al. 2001

Blood

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112

101

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“…In humans, cultivation of mast cells in serum-containing medium reduces the proliferation capacity of the mast cells because the serum-containing medium promotes growth of other cell types that englobe mast cells [18,19]. Lin et al [23] successfully established a culture method for generation of canine BMMCs using a serum-free medium.…”

Section: Discussionmentioning

confidence: 99%

“…Culture techniques have been developed to obtain a pure population of human mast cells derived from bone marrow [37], cord blood [36], peripheral blood [35], the fetal liver [19], skin [18], and the intestine [3]. Kambe et al [18] reported a successful culture system for proliferation of mature SMCs derived from dispersed human skin samples in a serum-free culture medium supplemented with recombinant human SCF.…”

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confidence: 99%

Cultivation and Characterization of Canine Skin-Derived Mast Cells

Kawarai

Masuda

Ohmori

et al. 2010

The Journal of Veterinary Medical Science

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ABSTRACT. 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 bluepositive metachromatic granules in the cytoplasm. The results of flow cytometric analysis for the presence of cell membrane c-Kit and Fc epsilon receptor I (FcRI) 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.KEY WORDS: bone marrow, canine, culture, mast cells, skin.J. Vet. Med. Sci. 72(2): 131-140, 2010 Allergic dermatitis in dogs is a complex of inflammatory skin diseases primarily mediated by type I hypersensitivity [8]. Cross-linking of the surface Fc epsilon receptor I (FcRI)-bound immunoglobulin E (IgE) with allergens results in the release of various mediators from mast cell granules, inducing type I hypersensitivity [5,6]. Studies using an experimental allergic model have revealed that cytokines and chemokines released from mast cells induce infiltration of leukocytes, leading to a late-phase reaction [42].In dogs, histopathological examination of the skin lesions of atopic dermatitis (AD) has revealed infiltration of mast cells, Langerhans cells, and lymphocytes in addition to a small number of eosinophils and neutrophils into the lesional skin [14]. The presence of allergen-specific IgEbearing mast cells in the dermis of allergic dogs has been shown [13], and both the number of mast cells and the histamine concentration in the dermis have been shown to correlate with clinical symptoms of AD [28]. As shown in the spontaneously developed AD in dogs, in a canine experimental model of AD, epicutaneous application of an allergen slurry has been shown to generate a penetration of allergen into the dermis, an elevated allergen-specific serum IgE, and localized pruritic dermatitis with typical histological changes by IgE-induced inf...

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“…Human cord blood-derived MCs (CBMCs) were cultured as described previously (25,26). Briefly, placentae were obtained within 45 min of delivery in adherence with ethics approval from the University of Alberta and Capital Health Region and with informed patient consent at Royal Alexandra Hospital, Edmonton, Alberta, Canada.…”

Section: Culturementioning

confidence: 99%

Protein Tyrosine Nitration of Aldolase in Mast Cells: A Plausible Pathway in Nitric Oxide-Mediated Regulation of Mast Cell Function

Sekar

Moon

Slupsky

et al. 2010

The Journal of Immunology

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An improved procedure for the development of human mast cells from dispersed fetal liver cells in serum-free culture medium

Cited by 32 publications

References 18 publications

Human skin–derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes

Human skin–derived mast cells can proliferate while retaining their characteristic functional and protease phenotypes

Cultivation and Characterization of Canine Skin-Derived Mast Cells

Protein Tyrosine Nitration of Aldolase in Mast Cells: A Plausible Pathway in Nitric Oxide-Mediated Regulation of Mast Cell Function

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