Co-culture of hepatocytes and fibroblasts by micropatterned immobilization of beta-galactose derivatives.

Photosensitive poly(allylamine) containing beta-galactose moieties in the side chain (LPAN(3)) was prepared by the reaction of poly(allylamine) with lactobionic acid and azidobenzoic acid. To create micropatterned surfaces, a LPAN(3)-coated poly(methyl methacrylate) (PMMA) substrate was irradiated with an ultraviolet lamp under a photomask. The presence of a LPAN(3) layer on the substrate was confirmed using an electron spectroscopy for chemical analysis. Micropatterned cell culture was carried out by seeding hepatocytes and/or fibroblasts on the substrate. As a result, hepatocytes and fibroblasts adhered only to the LPAN(3) and PMMA lane, respectively. Co-culture on the stripe patterned substrate was carried out by the first cell seeding of hepatocytes and subsequently by the second cell seeding of fibroblasts. The co-cultured cells produced extracellular matrix such as fibronectin, suggesting a biological function.