Ectopic transplantation of hepatocyte sheets fabricated with temperature-responsive culture dishes.

Aim: Hepatocyte transplantation to host livers by single cell suspension injection from the portal vein has been clinically successful in cases where the host liver architecture is intact. However, further investigation is still needed to achieve regeneration of the liver architecture when the host liver is destroyed, since single cell suspension injection often results in the formation of small hepatocyte colonies or islands. We show a hepatocyte transplantation strategy to ectopic sites. Methods: Primary hepatocytes isolated from green fluorescent protein (GFP)-transgenic rats were cultured on temperature-responsive culture dishes. After harvest as intact cell sheets, triple-layered cell sheets were transplanted over the superficial caudal epigastric artery and vein of athymic rats which had operation of 70% partial hepatectomy. Results: The transplanted hepatocytes were integrated to host tissue with a laminar cell arrangement at transplanted sites within one week after surgery. But the transplanted hepatocytes were hardly detected four weeks after transplantation, when the partially hepatectomized host liver was completely regenerated. GFP-positive bile duct-like tubes and functional blood vessels were observed. Conclusion: These results imply the usefulness of hepatocyte sheets in ectopic transplantation, as well as the need of trophic factors for hepatocyte survival.