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Journal of Histochemistry and Cytochemistry, Vol. 45, 1715-1722, Copyright © 1997 by The Histochemical Society, Inc.

TECHNICAL NOTE

Immunomagnetic Isolation of Rat Bone Marrow-derived and Peritoneal Mast Cells

Maria Celia Jamura, Ana Cristina G. Grodzkia, Andrea N. Morenoa, William D. Swaimb, Reuben P. Siraganianb, and Constance Oliverc
a Departamento de Biologia Celular, Universidade Federal do Paraná, Curitiba, PR, Brazil
b Laboratory of Immunology, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland
c Office of Naval Research, Arlington, Virginia

Correspondence to: Constance Oliver, Office of Naval Research, Code 335, 800 N. Quincy St., Arlington, VA 22217-5660.

Mast cells are difficult to purify from heterogeneous cell populations and to preserve, especially for pre-embedding immunostaining at the ultrastructural level. We have developed a technique that permits the isolation of a pure population of mast cells suitable for immunocytochemical studies. A rat mast cell-specific monoclonal antibody (MAb AA4) conjugated to tosylactivated Dynabeads 450 was used to immunomagnetically separate mast cells from rat bone marrow and peritoneal cell suspensions. Approximately 85% of the mast cells were recovered in the positive population that comprised virtually pure mast cells. After microwave fixation, morphological examination showed that the cells were intact and retained their ultrastructural detail. Mast cells in all stages of maturation were immunolabeled with a panel of antibodies after immunomagnetic separation. The combination of immunomagnetic separation followed by immunostaining should prove useful for the study of mast cell maturation and for the characterization of other specific cell types that are present in tissues in only limited numbers. (J Histochem Cytochem 45:1715-1722, 1997)

Key Words: mast cell, maturation, peritoneal cavity, bone marrow, immunomagnetic, immunocytochemistry, electron microscopy


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