Abstract
A method is described for the dissociation of guinea pig lung and mesentery into their component cells. The method comprises incubation of the tissues with the enzyme pronase in Ca++- and Mg++-free saline followed by mechanical dispersion and yields on average 3–8% free mast cells. These cells are morphologically intact and viable. They retain an active sensitization or can be passively sensitized. The levels of cAMP and cGMP in the whole cell suspension respond to catecholamine, histamine or cholinergic stimulation in a way similar to that observed in the corresponding undispersed tissue. Although the separation of guinea pig mast cells from other cells proved to be more difficult than the purification of rat mast cells, an inrichment of greater than 50% has been achieved. The mast cells isolated by this method proliferate in vitro and have been cultured successfully for several months.
Similar content being viewed by others
References
S.I. Said,Humoral Mediators in Bronchial Asthma: Implications for Therapy, inNew Directions in Asthma (Ed. M. Stein; Amer. Coll. Chest. Phys., Park Ridge, Illinois, 1975), pp. 261–273.
W.E. Brocklehurst,Many Facts, but Insufficient Knowledge: The Story of Asthma, J. Pharm. Pharmac.28, 361–368 (1976).
J.L. Mongar andH. O. Schild,Cellular Mechanisms in anaphylaxis, Physiol. Rev.42, 226–270 (1962).
M. Rocha e Silva (Sub-Ed.),Histamine and Anti-Histaminics, in O. Eiclar, A. Farah,Handbook of Experimental Pharmacology, Vol. 18/1, Springer, Berlin-Heidelberg-New York, 1966.
R. Keller,Tissue Mast Cells in Immune Reactions, Monographs in Allergy, 2, Karger, Basel and New York, 1966.
B. Uvnäs andI.-L. Thon,Isolation of ‘Biologically Intact’ Mast Cells, Exptl. Cell. Res.18, 512–520 (1959).
T.S.C. Orr andA.M.J.N. Blair,Potentiated Reagin Response to Egg Albumin and Conalbumin in Nippostrongylus Brasiliensis Infected Rats, Life Sci.8, 1073–1077 (1969).
W.M. Gold,Experimental Models of Asthma, inNew Directions in Asthma (Ed. M. Stein; Amer. Coll. Chest. Phys., Park Ridge, Illinois, 1975), pp. 241–249.
W.C. Bowman andC. Raper,Sympathomimetic Bronchodilators and Animal Models for Assessing their Potential Value in Asthma, J. Pharm. Pharmac.28, 369–374 (1976).
J.R. Carstairs, M.M. Dale andJ. L. Mongar,The Isolation and Some Properties of Guinea Pig Mesenteric Mast Cells, Exptl. Cell. Res. 2 (81), 156–162 (1973).
M.K. Bach, J.R. Brashler, K.J. Bloch andK.F. Austen,Studies on the Receptor Site for IgE Antibody on the Peritoneal Mast Cell of the Rat, in K.F. Austen, E.L. BeckerBiochemistry of the Acute Allergic Reaction, Blackwell, Oxford, 1971, pp. 65–89.
B.E.C. Banks, D.V. Banthorpe, D.M. Lamont, F.L. Pearce, K.A. Redding andC.A. Vernon,Dissociation of Sensory Ganglia from the Embryonic Chick by Pronase and Other Dispersing Agents, J. Embryol. exp. Morph.23, 519–530 (1970).
F.L. Pearce,Use of Porous Plastic Filters in the Preparation of Cell Suspensions, Microscopica acta (in press).
D.W. Aspray, A. Pitts andT.G. Pretlow II,Separation of Epithelial Cells from Suspensions of Cells from the Hamster Parotid Gland in an Isokinetic Density Gradient of Ficoll in Tissue Culture Medium, Anal. Biochem.66, 353–364 (1975).
T.G. Pretlow II andC.W. Boone,Separation of Mammalian Cells Using Programmed Gradient Sedimentation, Exp. Mol. Pathol. II, 139–152 (1969).
G. Poblete-Freundt, S. Glück andW. Schmutzler,Passive Sensitization with Homologous IgE and IgG 1 of Isolated Tissues and Mast Cells of the Guinea Pig (submitted).
A. Perini andI. Mota,The Production of IgE and IgG 1 Antibodies in Guinea Pigs Immunized with Antigen and Bacterial Lipopolysaccharides, Immunology25, 297–305 (1973).
A. G. Gilman,A Protein Binding Assay for Adenosine 3′,5′ Cyclic Monophosphate, Proc. Nat. Acad. Sci. (Wash.)67, 305–312 (1970).
V. Dinnendahl,A Rapid and Simple Procedure for the Determination of Guanosine 3′,5′-Monophosphate by Use of the Protein-Binding Method, Arch. Pharmacol.284, 55–61 (1974).
F. Murad, V. Manganiello andM. Vaughan,A Simple, Sensitive Protein-Binding Assay for Guanosine 3′,5′-Monophosphate, Proc. Nat. Acad. Sci. (Wash.)68, 736–739 (1971).
O.H. Lowry, N.J. Rosebrough, A.L. Farr andJ.R. Randall,Protein Measurement with the Folin Phenol Reagent, J. Biol. Chem.193, 265–275 (1951).
W. Schmutzler, G. Poblete-Freundt andR. Derwall,Studies on the Role of cAMP in the Regulation of the Anaphylactic Histamine Release from Guinea Pig Lung, Agents and Actions3, 192–193 (1973).
A.A. Mathé, L. Volicer andS.K. Puri,Effect of Anaphylaxis and Histamine, Pyrilamine and Burimamide on Levels of Cyclic AMP and Cyclic GMP in Guinea Pig Lung, Res. Comm. Chem. Path. Pharmacol.8, 635–651 (1974).
J. Stoner, V.C. Manganiello andM. Vaughan,Guanosine Cyclic 3′,5′-Monophosphate and Guanylate Cyclase Activity in Guinea Pig Lung: Effects of Acetylcholine and Cholinesterase inhibitors, Mol. Pharmacol.10, 155–161 (1974).
L.O. Boreus andN. Chakravarty,The Histamine Content of Guinea Pig Mast Cells, Experientia (Basel)16, 192 (1960).
N.S. Taichman,Ultrastructure of Guinea Pig Mast Cells, J. Ultrastructure Res.32, 284–292 (1970).
H. Behrendt, Ch. Stang-Voss, U. Blum, F. L. Pearce, G. Poblete-Freundt andW. Schmutziler,The Ultrastructure of Isolated Guinea Pig Mast Cells (in preparation).
H. Ginsburg,The in vitro Differentiation and Culture of Normal Mast Cells from the Mouse Thymus, Ann. N.Y. Acad. Sci.103, 20–39 (1963).
Ch. Stang-Voss, H. Behrendt, U. Blum, B. Fischer, F.L. Pearce, G. Poblete-Freundt andW. Schmutzler,Studies on Cultured Guinea Pig Mast Cells (in preparation).
W. Schmutzler, H. Behrendt, U. Blum, F.L. Pearce, G. Poblete-Freundt andCh. Stang-Voss,Histamine Release from Isolated Guinea Pig Mast Cells by Anaphylatoxin, Transact. Coll. Int. Allergol., 1976, Monographs in Allergy (in press).
H. Behrendt, U. Blum, F.L. Pearce, G. Poblete-Freundt, P. Pult, W. Schmutzler andCh. Stang-Voss,Zur Isolierung von Mastzellen des Meerschweinchens, Anat. Anz. Erg. H. (in Press).
P. Pult, A. Krämer andW. Schmutzler,Pharmacological Studies in Isolated Mast Cells from Human Nasal Mucosa, Allergologia et Immunopathologia4, 195 (1976).
Author information
Authors and Affiliations
Additional information
Parts of this work have been presented to the Spring Meeting of the German Pharmacological Society, Mainz, March 1976, to the Anatomical Society, Rostock, April 1976, and to the Annual Meeting of the European Branch of the Histamine Club, Paris, May 1976.
Rights and permissions
About this article
Cite this article
Pearce, F.L., Behrendt, H., Blum, U. et al. Isolation and study of functional mast cells from lung and mesentery of the guinea pig. Agents and Actions 7, 45–56 (1977). https://doi.org/10.1007/BF01964880
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01964880