Immunomagnetic isolation of rat bone marrow-derived and peritoneal mast cells

Maria Celia Jamur, Ana Grodzki, Andrea N. Moreno, William D. Swaim, Reuben P. Siraganian, Constance Oliver

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1715-1722
Number of pages8
JournalJournal of Histochemistry and Cytochemistry
Volume45
Issue number12
DOIs
StatePublished - Jan 1 1997
Externally publishedYes

Keywords

  • Bone marrow
  • Electron microscopy
  • Immunocytochemistry
  • Immunomagnetic
  • Mast cell
  • Maturation
  • Peritoneal cavity

ASJC Scopus subject areas

  • Anatomy
  • Histology

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