A LINE OF RAT OVARIAN SURFACE EPITHELIUM PROVIDES A CONTINUOUS SOURCEOF COMPLEX EXTRACELLULAR-MATRIX

A spontaneously immortalized, yet non-tumorigenic rat ovarian surface epithelial (ROSE 199) cell line, deposits large amounts of extracellul ar matrix (ECM) in response to crowding. The characteristics and compo nents of ROSE 199-derived cell-free ECM were compared after three diff erent preparative techniques: treatment with 20 mM ammonium hydroxide, with 1% sodium deoxycholate, or by repeated freeze-thaws. The ECMs we re analyzed by histochemistry, immunofluorescence, electron microscopy , and Western immunoblotting. Components of ROSE 199 ECM included lami nin, fibronectin, and collagen types I and III. Even though ROSE 199 i s an epithelial cell line, striated collagen fibers formed a major par t of its matrix. Thus, ROSE 199 matrix consists of both basement membr ane and stromal matrix components. This matrix supported the adhesion, spreading, and growth of several cell types without altering their mo rphology or growth pattern, and enhanced the attachment of some cell t ypes that spread on plastic only with difficulty. Immunofluorescence, electron microscopy, and dry weight determinations indicated that a gr eater proportion of matrix was retained in preparations obtained by am monium hydroxide or freeze thaw techniques than after sodium deoxychol ate treatment. Ammonium hydroxide and freeze-thaw treated matrices wer e also superior to sodium deoxycholate preparations as evidenced by en hanced initial cellular adhesion and spreading compared to cells plate d on plastic. Residual nuclear material did not seem to affect the bio logical activity of this matrix. ROSE 199 extracellular matrix provide s a novel, complex substratum for cell culture and for studies of matr ix functions and synthesis.