Distribution of Type I Collagen and Basement Membrane Proteins in the Cultures of Cloned Epithelial Liver Cells

摘要

References(36) Cited-By(1) Cultures of a cloned epithelial cell lines derived from rat liver that synthesizes α-fetoprotein were used. Time courses of the appearance and distribution of types I and IV collagen, laminin and fibronectin in these cultures were assayed by the unlabeled peroxidase anti-peroxidase technique. Fibrillogenesis was monitored by the conventional phosphotungstic acid aniline-blue staining method for collagen fibrils.In the initial stage of cell proliferation, the antigenecities of these four matrix proteins were found only in the interior of most cells. Later, laminin and fibronectin began to appear in the intercellular spaces, although types I and IV collagens still were confined within the cells. With the onset of fibrillo-genesis, extracellular deposition of types I and IV collagen began together with a relative decrease in the intensity of cellular staining. As fibrillogenesis progressed, these four matrix proteins gave more and more intense immuno-cytochemical reactions in the extracellular spaces, developing into the charac-teristic circular pattern of distribution. This process usually stopped when fibrillogenesis reached what appeared to be a static state. Immunocytochemical staining for the type I collagen of the preparations, then phosphotungstic acid aniline blue staining showed that two types of staining coincide. In contrast, none of three other matrix proteins showed complete codistribution with aniline blue positive fibrils. They were, however, located close to each other.Our results show that cultured epithelial liver cells produce these matrix proteins from a time early in culture, and that the extracellular depositions of these proteins take place successively with some interrelation.