title: Novel approach for achieving double-layered cell sheets co-culture: overlaying endothelial cell sheets onto monolayer hepatocytes utilizing temperature-responsive culture dishes.
abstract: Confluent human aortic endothelial cells (HAECs) cultured on thermo-responsive culture dish grafted with poly (N-isopropylacrylamide) were recovered as a contiguous cell sheet. The double-layered co-culture was achieved by placing the recovered HAEC sheet onto the rat hepatocyte layer directly. The double-layered structure of HAEC and hepatocytes remained in tight contact during culture. Hepatocytes in the layered co-culture system with the HAEC sheet maintained the differentiated cell shape and the albumin expression for over 41 days of culture, whereas the functions disappeared within 10 days of culture in control hepatocytes without the HAEC sheet. The layered co-culture of hepatocytes and the HAEC sheets, which allows for the expression of differentiated functions of hepatocyte continuously, such as liver lobule, offers a major advancement in liver tissue engineering.
authors.affiliation: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
authors.forename: Masami
authors.initials: M
authors.lastname: Harimoto
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yamato
authors.forename: Motohiro
authors.initials: M
authors.lastname: Hirose
authors.forename: Chie
authors.initials: C
authors.lastname: Takahashi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Isoi
authors.forename: Akihiko
authors.initials: A
authors.lastname: Kikuchi
authors.forename: Teruo
authors.initials: T
authors.lastname: Okano
mesh_terms: D018920:Coculture Techniques; D004730:Endothelium, Vascular / Q000166:cytology*; D004864:Equipment and Supplies; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D008856:Microscopy, Fluorescence; D013696:Temperature