Isolation, Culture, and Characterization of Human Intestinal Smooth Muscle Cells

In vitro models utilizing human mesenchymal cells isolated from normal and pathological tissue have proven very useful for studies of wound repair and fibrosis. We have been studying the pathogenesis of intestinal fibrosis and, over the course of 20 yr, have refined techniques for the isolation, culture, and characterization of smooth muscle cells of the human intestinal muscularis propria (1 ). These methodologies have been adapted for isolation of similar cell types from liver, gallbladder, and blood vessels (2 ). In this chapter, we describe techniques for the isolation and culture of these cells (procedure A), for the quantitation of procollagen secretion (procedure B), and for the screening of isolates for the expression of smooth muscle-specific cytoskeletal proteins (procedure C).