IDENTIFICATION OF PURE AND MIXED BASOPHIL COLONIES IN CULTURE OF HUMAN PERIPHERAL-BLOOD AND MARROW-CELLS

  • 1 January 1984
    • journal article
    • research article
    • Vol. 64  (1) , 78-83
Abstract
A colony assay system is presented that allows in situ identification of human basophil/mast cell (basophil) colonies. In methylcellulose culture, in the presence of phytohemagglutinin-leukocyte conditioned media (PHA-LCM), human peripheral blood and bone marrow cells form colonies that can be distinguished by their unique morphological characteristics. Pure basophil colonies are diffuse, small colonies containing small, round, highly refractile cells. These characteristics of the constituent cells show that a significant number of basophils are found in combination with eosinophils. The mixed eosinophil/basophil colonies have the distinctive elements of pure eosinophil and pure basophil colonies. Usually, these are diffuse colonies with compact clusters of slighly larger, darker-appearing cells. Colonies were found that contained basophils and neutrophils/monocytes, but this type could not be consistently identified by in situ morphology. Cytochemical analysis confirmed the metachromatic nature of the granules in the basophils. The presence of IgE receptors on the cells was documented by indirect immunofluorescent staining after passive sensitization with purified human IgE. Peripheral blood cells from 6 healthy volunteers formed 5.7 .+-. 1.0 (mean .+-. SEM [standard error of the mean]) pure colonies in 2 .times. 105 cells. Cultures of bone marrow cells from patients with various types of anemia had 9.0 .+-. 1.5 colonies in 105 cells. This is the 1st description of a colony assay system for in situ identification of a pure population of basophilic granulocytes.