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Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies
Published on: December 26, 2012
Cytoplasmic bar-like structures of alveolar type II cells: an ultrastructural study in freshly isolated cells from
Abstract:
Bar-like structures are tubular cytoplasmic inclusions found in situ in pulmonary epithelial type II cells of several animal species. The physiological significance and mode of formation of these inclusions are not fully established. In this paper, we describe bar-like structures as found in freshly isolated type II cells from rat lungs. Pulmonary cells were dissociated from the tissue with elastase and separated on a discontinuous density gradient of Percoll. The complete isolation procedure yielded 17 X 10(6) type II cells per animal (purity = 80%). Either from the crude cell suspensions or the purified preparations, only a small fraction of the type II cell population harbored the inclusions (less than 5%). It is shown that the bounding membranes of the bar-like structures can derive from the endoplasmic reticulum, the nuclear membrane, or the Golgi apparatus. Occasional connections with lamellar bodies were observed, and different levels of complexity in the bar-like structures were also found. The apparent rigid conformation and the orientation of the bar-like structures were taken as evidence for a role of the cytoskeleton in their formation. Because the inclusions do not appear to be new organelles or cellular structures performing a specific function, we propose that their formation may be a transient and limited cellular event in normal cells. However, the stabilization and the generation of the osmiophilic structures, as well as their overproduction, may reflect alterations of the normal physiology of the type II cells.
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