title: Engineering of an hepatic organoid to develop liver assist devices.
abstract: Cell-based technologies to support/restore liver function represent one of the most promising opportunities in the treatment of acute liver failure. However, the understanding of the constituent cell types that interact to achieve liver-specific structure and function has not been achieved in the development of liver assist devices (LADs). Here we show that hepatocytes migrated toward and adhered and formed sinusoids-like structures in conjunction with liver nonparenchymal cells, and that this liver organoid formed sophisticated tissue after 7 days in an implanted LAD in rodents. Hepatocytes only or in combination with human nonparenchymal liver cell lines (endothelial, cholangiocytes, and stellate cells) were cultured in Matrigel. Ultrastructural analysis showed that the hepatocyte-decorated endothelial vascular structures resemble in vivo sinusoids containing plate-like structures, bile canaliculi, and lumen. The sinusoid-like structures retained albumin secretion and drug metabolism capabilities. In addition, LADs containing cocultures of human liver nonparenchymal cells were transplanted in animals for a week; the liver tissue formed sophisticated structures resembling the liver. These results demonstrate the importance of nonparenchymal cells in the cellular composition of LADs. The novelty of the culture's sinusoid-like organization and function strongly support the integration of liver nonparenchymal units into hepatocyte coculture-based LADs as a potential destination therapy for liver failure.
authors.affiliation: Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama, Japan. alejandro.sotogutierrez@chp.edu
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutierrez
authors.forename: Nalú
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yagi
authors.forename: Yakoov
authors.initials: Y
authors.lastname: Nahmias
authors.forename: Martin L
authors.initials: ML
authors.lastname: Yarmush
authors.forename: Naoya
authors.initials: N
authors.lastname: Kobayashi
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D018920:Coculture Techniques; D006498:Hepatectomy; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D020007:Implants, Experimental; D008099:Liver / Q000502:physiology* / Q000601:surgery; D008111:Liver Function Tests; D019164:Liver, Artificial; D051379:Mice; D008807:Mice, Inbred BALB C; D009940:Organoids / Q000502:physiology* / Q000648:ultrastructure; D023822:Tissue Engineering / Q000379:methods*