Phenotypic evaluation of cultured human mast and basophilic cells and of normal human skin mast cells

Abstract

In order to evaluate various markers for human mast cells, two human mast/basophilic cell lines (HMC-1/KU812), cultured mast cells from the peripheral blood monocytic fraction and peripheral blood monocytes were compared with mast cells in tissue sections from normal skin, using histochemistry, enzyme histochemistry and immunohistochemistry. All reagents stained normal skin mast cells, with toluidine blue, tryptase reactivity and antibodies against the Fc epsilon RI and the stem cell factor receptor (c-kit) being most active. The cell lines and mast cells cultured from peripheral blood were negative for avidin, safranin and chymase, strongly positive for c-kit and variably reactive with all other reagents. All antibodies except AA1 against tryptase also stained one or several epidermal and dermal cell types or blood monocytes. Histochemical stains (toluidine blue, avidin) and reagents for the enzymes tryptase and chymase are thus specific markers for mast cells. The frequent reactivity of antibodies against mast cells with other cell types indicates interesting functional and ontogenetic relationships between these cells.

Synthesis, Storage, and Release of Vascular Endothelial Growth Factor/Vascular Permeability Factor (VEGF/VPF) by Human Mast Cells: Implications for the Biological Significance of VEGF206

“…Therefore, it is reasonable to predict that mast cells may enhance synthesis of VEGF in vivo when activated under inflammatory conditions. Since HMC-1 cells only variably express the highaffinity receptor for IgE (Hamann et al, 1994;Nilsson et al, 1994), an IgE-like activation of these cells was achieved by triggering calcium influx, combined with PKC activation (Sagi-Eisenberg, 1993). HMC-1 cells activated in this way start de novo synthesis of various mediators, such as IL-1␤, IL-6, IL-8, and bFGF (Grabbe et al, 1995;Qu et al, 1995).…”

“…The bulk of experiments performed in this study was done with the cell line HMC-1, representing the only established human cell line exhibiting a phenotype similar to that of normal human tissue mast cells (Hamann et al, 1994;Nilsson et al, 1994). Since VEGF is overexpressed in several transformed cell lines (White et al, 1995), we cannot exclude the possibility that the expression of VEGF in HMC-1 cells is less a mast cell-associated than a tumor-specific characteristic.…”

“…HMC-1 cells (kindly provided by Dr. Butterfield, Rochester, MN), which are immature human leukemic mast cells (Butterfield et al, 1988;Hamann et al, 1994), were cultured in Iscove's medium (Seromed, Berlin, Germany), supplemented with 10% fetal calf serum (FCS) (Seromed), 10 M monothiolglycerol (Sigma Chemical, Deisenhofen, Germany), and antibiotics (streptomycin and penicillin). Stimulation of HMC-1 cells was performed in 24-multiwell plates (Falcon, Lincoln Park, NJ), as previously described (Mö ller et al, 1993).…”

Expression of SCF splice variants in human melanocytes and melanoma cell lines: potential prognostic implications

“…Immunocytochemistry was performed and evaluated on cytocentrifuge preparations of cells stained with the APAAP technique, as previously described (Hamann et al, 1994). Three different antibodies against SCF were used, a monoclonal antibody against SCF purchased from Genzyme (Cambridge, MA, USA), a mouse monoclonal antibody (HKL-12), and a rabbit polyclonal antibody (7278) , both kindly donated by M Brockhaus, Basel, Switzerland.…”

Establishment and Characterization of a Murine Mast Cell Line Derived from NC/Nga Mice

“…After 3 weeks of culture, over 90% of the nonadherent cells are identifiable as mast cells based on their characteristic morphological appearance, metachromasia of the granules with the toluidine blue stain, positive Alcian blue staining, and surface expression of the high affinity IgE receptor (FcεRI) and c-kit. In contrast to mature-type mast cells such as mucosal-type mast cells which reside in the intestinal mucosa, and connective tissue-type mast cells found in the skin and peritoneal cavity, BMMCs are negative for safranin staining and considered to be of an immature phenotype [3, 4]. …”