Growth of macrophage colonies from normal canine peripheral blood: morphological, cytochemical and functional...

Canine macrophage colonies were grown at high colony-forming efficiencies (average of 4.5 macrophage colonies/10(4) mononuclear cells plated) from gradient-separated peripheral blood mononuclear cells. The colonies were first observed on day 5 of the culture period, reached maximum numbers between days 10-14, and differed kinetically from CFU-GM colonies. Colony cells had typical macrophage morphology at the cellular and ultrastructural levels, were nonspecific-esterase positive, specific-esterase negative, and were actively phagocytic. Colony growth in semisolid cultures and 3H-TdR incorporation in liquid cultures occurred following stimulation with rabbit anti-canine immunoglobulin antisera (anti-Ig), anti-bovine serum albumin and normal rabbit serum. Addition of 2-mercaptoethanol (2-Me) to stimulated cultures significantly enhanced the proliferative response. A maximal response was obtained using anti-IgM and 2-ME. Preincubation of anti-Ig with goat anti-rabbit IgG or purified canine immunoglobulin in the presence or absence of 2-ME significantly reduced the proliferative response, suggesting the presence of both specific and nonspecific components of stimulation. The growth of canine macrophage colonies from peripheral blood provides a method for non-invasive, sequential and kinetic studies of macrophage progenitor cells in large animals.