Coated antireceptor antibody as an extracellular matrix for liver tissue engineering

Differences of hepatocyte attachment and morphology of adherent cells between anti-asialoglycoprotein receptors (α-ASGPR) and poly (N-p- vinylbenzyl-O-β-D-galactopyranosyl-[1 → 4]-D-gluconamide) (PVLA) coated surfaces were investigated for a new application of antibody as an extracellular matrix. It was found that about 70% of mouse primary hepatocytes attached onto the α-ASGPR-coated plates within 2 h. The hepatocyte attachment was inhibited with pretreatment of soluble α-ASGPR or PVLA. Also, the number of the adherent hepatocytes drastically decreased if treated with papain as an endopeptidase but not with EDTA. This was in contrast with the hepatocytes attached onto PVLA-coated plates, suggesting that cell attachment through specific interaction between the ASGPR and α- ASGPR was Ca²⁺ ions-independent. The adherent hepatocytes exhibited quite different morphology with the coating density of the α-ASGPR. The morphology of adherent hepatocytes for a high coating density (10 μg/ml) was observed as spherical shapes whereas the morphology of hepatocytes for a low coating density (0.1 μg/ml) was observed as fiat ones. Furthermore, when cellular function was examined for hepatocytes with different morphology, it was found that DNA synthesis of fiat hepatocytes for the lower coating densities was higher than that of round hepatocytes for the higher coating densities in the presence of epidermal growth factor.