Figure 1

Establishment and optimisation of the liver perfusion culture system.

(a), Diagram of the ex vivo organ perfusion culture circuit. (b), Photograph of isolated liver placement by our hanging method in the organ chamber. (c), Assessments of ALT activity (left), albumin synthesis (centre) and bile production (right) during liver perfusion culture at 22°C with 5.0 × 10¹¹ cells/L erythrocytes. The coloured lines represent temperatures of 37°C (black), 33°C (green), 22°C (red), 10°C (purple) and 4°C (blue). (d), Photographs (top) and histological images (middle and bottom) of the cultured livers at different temperatures. Higher-magnification images are shown in the boxed area (bottom). Scale bars, 100 μm. (e), Assessments of ALT activity (left), albumin synthesis (centre left), bile production (centre right) and urea synthesis (right) during liver perfusion culture at 22°C with/without erythrocytes. These data represent erythrocyte concentrations of 0.5 × 10¹¹ cells/L (green), 2.0 × 10¹¹ cells/L (orange), with 5.0 × 10¹¹ cells/L (red) and no erythrocytes (blue). (f), 3D images of sinusoidal structure in natural (top column) and cultured livers after 48 hr of perfusion at 22°C with/without erythrocyte (middle column and bottom column). The sagittal section (top) and horizontal section (bottom) are shown in a hepatic lobule between the central vein (CV) and the portal vein (PV). The vascular structure is represented by FITC (green) and dead cells (red) are indicated by the propidium iodide staining. All sections are counterstained with Hoechst 33342 (blue). Scale bars: 100 μm. (g), The numbers of total cells (white) and dead cells (black) in the natural (left) and cultured livers after 48 hrs at 22°C with/without erythrocytes (centre and right). These cells are counted in the optional areas evenly between CV and PV in a range of 100 μm (wide) × 200 μm (depth) × 100 μm (height). (h), Histological analysis of natural (left) and cultured livers after 48 hrs at 22°C with/without erythrocytes (centre and right). Higher-magnification images are shown in the boxed area (bottom). Scale bars, 100 μm.