Adult rat hepatocytes efficiently attach to intact connective tissue fibers prepared from normal rat liver; this material has been given the name of %omatrix". The cells remain alive and differentiated for at least 4 months in culture. The liver biomatrix contains both collagen and noncollagenous glycoproteins. Heretofore, the functions of the different components of the biomatrix in facilitating cell adhesion and promoting survival and differentiation of rat hepatocytes have not been investigated. We now report on an aggregate of glycoproteins which were extracted with dilute acetic acid from liver biomatriar and which facilitate hepatocyte adhesion to plastic dishes and collagen. One protein in the extract, with a molecular weight larger than 300,000, was selectively immunoprecipitated with an antibody prepared against the extract and immunoadsorbed to glutaraldehyde-cross-linked rat serum. The antibody did not cross-react with cold-insoluble globulin and selectively inhibited cell attachment. The protein of molecular weight of 300,000 appears to be specific for liver, since similar extracts prepared from rat lung, kidney, and heart biomatrices did not cross-react with the antibody. Furthermore, the latter extracts did not facilitate cell attachment above values obtained with plain culture dishes. The results suggest that glycoproteins in the biomatrix, which are antigenically distinct from fibronectin, can mediate attachment of rat hepatocytes to plastic dishes and collagen.