title: Consideration of a Safe Protocol for Hepatocyte Transplantation Using Infantile Pigs.
abstract: Hepatocyte transplantation is hoped to be an alternative treatment to certain cases of liver transplantation. The most promising indication is for congenital metabolic diseases, especially in infants. To establish a safe protocol for hepatocyte transplantation into infants, we examined physiological changes during treatment using infantile pigs. Recipient pigs (domestic, crossbred with Large-Yorkshire, Landrace, and Duroc; 2.5 kg; 7-14 days old) were anesthetized with isoflurane; a midline incision of minimum length provided access to the superior mesenteric vein. A double lumen catheter was inserted through the vein to within 1 cm of the hepatic portal region. Physiological parameters such as heart rate, systemic blood pressure, and portal pressure were monitored. Cryopreserved porcine hepatocytes isolated from the same strain were suspended in physiological saline and transfused through the catheter. In experiments requiring tracing, cells were stained with fluorescent dye prior to transfusion. Recipient pigs were kept for 1 day and sacrificed for histological liver examination. After preliminary experiments, the optimized number and concentration for hepatocyte transplantation were determined to be 1 × 108cells/kg and 1 × 107cells/ml. The cell suspension was transfused at a rate of 0.67 ml/min. No marked anomaly of physiological parameters was observed, whereas light tachycardia occurred in preliminary trials when the transfusion rate was faster than our standard protocol. All five pigs transfused with the established method recovered from anesthesia and survived with good vital signs. Histology revealed that the transfused hepatocytes were integrated in the hepatic tissue.
authors.affiliation: Clinical Research Center, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Shin
authors.initials: S
authors.lastname: Enosawa
authors.affiliation: Clinical Research Center, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Wenji
authors.initials: W
authors.lastname: Yuan
authors.affiliation: Clinical Research Center, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Masaharu
authors.initials: M
authors.lastname: Douzen
authors.affiliation: † Department of Pathology, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Atsuko
authors.initials: A
authors.lastname: Nakazawa
authors.affiliation: ‡ Institute of Technology and Science, The University of Tokushima , Tokushima , Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Omasa
authors.affiliation: § Division of Transplantation, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Akinari
authors.initials: A
authors.lastname: Fukuda
authors.affiliation: § Division of Transplantation, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Seisuke
authors.initials: S
authors.lastname: Sakamoto
authors.affiliation: § Division of Transplantation, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Takanobu
authors.initials: T
authors.lastname: Shigeta
authors.affiliation: § Division of Transplantation, National Center for Child Health and Development , Tokyo , Japan.
authors.forename: Mureo
authors.initials: M
authors.lastname: Kasahara
keywords: Hepatocyte transplantation; Infantile pigs; Safe protocol