Abstract
Abstract Introduction: Hepatocyte cellular transplantation, a potential alternative to liver transplantation, has been successfully applied in animal models and in clinical studies as a bridge to transplantation. This approach is challenged by limited donor hepatocytes and by the inability to maintain the long-term survival and expansion of hepatocytes in vitro. We propose that the in vitro expansion and eventual therapeutic delivery of liver cells would be enhanced by co-culture with endothelial cells in active angiogenesis. Methods: Human fetal liver cells, enriched for hepatocyte progenitor cells (HPC) by FACS sorting, were co-cultured with human umbilical vein endothelial cells (HUVEC) either in a monolayer or in a 3D fibrin gel system (to induce angiogenesis). Angiogenesis in the 3D system was quantified by the Simple PCI software. HPC, transduced with the human albumin promoter-directed DsRed were tracked by microscopy and proliferation was measured by BrdU labeling. Alpha-fetoprotein and albumin was measured by ELISA. The association of HPC and HUVEC was examined by confocal microscopy. Results: Angiogenesis occurred within 16 days in 3D cultures. HPC survived longer in 3D HUVEC cocultures [> 60 days] than in 2D cocultures [26 ± 7 days] or when cultured alone [28 ± 6 days]. BrdU confirmed HPC proliferation in the 3D system and albumin production was greater in 3D cocultures [177.7 μg/mL/day] than in the 2D cocultures [30 μg/mL/day] or in HPC alone cultures [73.6 μg/mL/day]. Conclusions: Survival and expansion of liver HPC was greatly enhanced when in association of EC in active angiogenesis.
Published Version
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