Hepatocyte transplantation and sample preparation.

<p>(A) Hepatocyte transplantation. After retrorsine treatment, the parenchyma of the liver became slightly hard and the surface was irregular. After resection of the left lobe and the left side of the middle lobe, hepatocytes were transplanted directly into the liver. The suspended cells spread rapidly according to the blood flow and produced a demarcation line, which disappeared immediately after injection. (B) Isolated hepatocytes. Trypan blue staining identified the viable hepatocytes, which were negative for 7-AAD (red stain) and expressed GFP. GFP-positive hepatocytes lose GFP expression immediately after cell death. The isolated hepatocytes cells were not completely separated. (C) Sample collection. Two weeks after cell transplantation, the remnant liver grew enlarged and swollen. (Upper arm) A small piece of the liver was obtained from the right edge of the middle lobe before perfusion from the portal vein, and then cut into smaller pieces for PCR analysis. The larger part of the liver was fixed with 2% paraformaldehyde for microscopic observation. (Lower arm) The liver was perfused with 2% paraformaldehyde and removed. The right part of the middle lobe was cut into three pieces and further processed for microscopic observation.