Summary
The skin is the major site on anaphylaxis, and cutaneous mast cells have an important role in its reactions. The isolation and purification of rat cutaneous mast cells are described here. Rat abdominal skin was digested with collagenase and hyaluronidase, and centrifuged with Percoll. The buoyant density of cutaneous mast cells was high, and relatively pure mast cells were obtained. The purity of cutaneous mast cells was 7.4%±2.4% before and 50.0%±6.4% after Percoll density centrifugation; peritoneal mast cells revealed 5.8%±1.3% purity before and 61.0%±10.6% purity after the same procedure. The isolated cutaneous cells released 21.3%±3.8% histamine and the peritoneal mast cells released 55.5%±3.8% histamine upon stimulation with 10 μg/ml compound 48/80. These findings suggest that there are functional subsets of connective tissue mast cells.
Similar content being viewed by others
References
Barrett KE, Ali H, Pearce FL (1985) Studies on histamine secretion from enzymically dispersed cutaneous mast cells of the rat. J Invest Dermatol 84:22–25
Benyon RC, Lowman MA, Church MK (1987) Human skin mast cells: their dispersion, purification, and secretory characterization. J Immunol 138:861–867
Coutts SM, Nehring RE Jr, Jariwala NU (1980) Purification of rat peritoneal mast cells: occupation of IgE receptors by IgE prevents loss of the receptors. J Immunol 124:2309–2315
Enerbäck L (1981) The gut mucosal mast cell. Monogr Allergy 17:222
Enerbäck L, Sevensson I (1980) Isolation of rat peritoneal mast cells by centrifugation on density gradients of Percoll. J Immunol Methods 39:135–145
Gibson S, Miller HRP (1986) Mast cell subsets in the rat distinguished immunohistochemically by their content of serine proteinases. Immunology 58:101–104
Hachisuka H, Nomura H, Sasai Y, Shiotsuki K, Yokoyama MM (1986) Changes of intracellular fluidity during keratinocyte differentiation measured by fluorescence polarization. Cell Tissue Res 246:557–560
Hachisuka H, Nomura H, Sakamoto F, Mori O, Okubo K, Sasai Y (1988) Effect of antianaphylactic agents on substance-P induced histamine release from rat peritoneal mast cells. Arch Dermatol Res 280:158–167
Heiman AS, Crews FT (1985) Characterization of the effects of phorbol esters on rat mast cell secretion. J Immunol 134:548–555
Ishizaka T, König W, Kurata M, Mauser L, Ishizaka K (1975) Immunologic properties of mast cells from rats infected with Nippostrongylus brasiliensis. J Immunol 115:1078–1083
Koda A, Nagai H, Watanabe S, Yanagihara Y, Sakamoto K (1976) Inhibition of hypersensitivity reactions by a new drug, N(3′,4′-dimethoxycinnamoyl) anthranilic acid (N-5′). J Allergy Clin Immunol 57:396–407
Kusner EJ, Dubnick B, Herzig DJ (1973) The inhibition by disodium cromoglycate in vitro of anaphylactically induced histamine release from rat peritoneal mast cells. J Pharmacol Exp Ther 184:41–46
Martin U, Römer D (1978) The pharmacological properties on a new, orally active antianaphylactic compound: ketotifen, a benzocycloheptathiophene. Arzneimittelforsch 28:770–782
Newlands GF, Huntley JF, Miller HR (1984) Concomitant detection of mucosal mast cells and eosinophils in the intestines of normal and Nippostrongylus-immune rats. A reevaluation of histochemical and immunocytochemical techniques. Histochemistry 81:585–589
Sasai Y, Hachisuka H, Mori O, Nomura H (1984) Separation of keratinocytes by density gradient centrifugation for DNA cytofluorometry. Histochemistry 80:133–136
Schleimer RP, Macglashan DW Jr, Peters SP, Pinckard RN, Adkinson NF Jr, Lichtenstein LM (1986) Characterization of inflammatory mediator release from purified human lung mast cells. Am Rev Respir Dis 133:614–617
Shanahan F, Denburg JA, Fox J, Bienenstock J, Befus D (1985) Mast cell heterogeneity: effects of neuroenteric peptides on histamine release. J Immunol 135:1331–1337
Sullivan TJ, Parker KL, Stenson W, Parker CW (1975) Modulation of cyclic AMP in purified rat mast cells. J Immunol 114:1473–1479
Tsuruta Y, Kohashi K, Ohkura Y (1978) Determination of histamine in plasma by high-speed liquid chromatography. J Chromatogr 146:490–493
Uvnäs B, Thon I-L (1959) Isolation of “biologically intact” mast cells. Exp Cell Res 18:512–520
Yurt RW, Leid RW Jr, Spragg J, Austen KF (1977) Immunologic release of heparin from purified rat peritoneal mast cells. J Immunol 118:1201–1207
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hachisuka, H., Kusuhara, M., Higuchi, M. et al. Purification of rat cutaneous mast cells with Percoll density centrifugation. Arch Dermatol Res 280, 358–362 (1988). https://doi.org/10.1007/BF00426614
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00426614