ラクトース修飾絹フィブロイン基材上における初代培養ラット肝細胞のスフェロイド形成と維持

Abstract

We examined the formation and maintenance of spherical multicellular aggregates (spheroids) of rat primary hepatocytes on dishes coated with lactose-silk fibroin conjugates bearing galactose residues (Lac-CY-SF) as substrate materials. Rat hepatocytes that had attached onto the conjugate-coated dishes were subsequently cultured in a medium supplemented with epidermal growth factor (EGF) and insulin. After the rat hepatocytes extended flat on the conjugate-coated dishes in 2 days of culture, these hepatocytes formed spheroids about 100 to 300 μm in diameter at day 4. After 6 days of culture the spheroids were maintained without any obvious change in size on the conjugate-coated dishes. However, the detachment of the spheroids from the conjugate-coated dishes began from day 8, and the spheroids clearly shrank at day 10. These results suggested that hepatocyte spheroid formation was induced on the Lac-CY-SF conjugate-coated dishes in the presence of EGF and insulin, but the conjugate-coated dishes were not capable of keeping the spheroids for more than 5 days.