Isolation, Transplantation, and Long-Term Noninvasive Tracking of DiD-Labeled EpCAM+ Human Fetal Hepatic Progenitors in Mouse Livers.

Abstract

Chronic liver disease (CLD) is a progressive condition characterized by the deterioration of liver structure and function, resulting from persistent injury and inflammation. Liver cell therapy has emerged as a promising alternative bridging strategy for patients waiting for the availability of a suitable donor liver for transplantation. Fetal human hepatic progenitor cells (fHPCs) hold great potential as a source for liver regeneration and restoration of liver function in individuals with CLD. A key challenge in liver cell therapy lies in the ability to effectively track transplanted donor cells, monitoring their homing, repopulation, and functional integration into the recipient's liver.This protocol outlines a comprehensive methodology for isolating fHPCs, enrichment of EpCAM positive cells, and labeling them with DiD dye. It also details the procedure for inducing liver fibrosis in SCID mice, transplanting the donor fHPCs, and conducting noninvasive, long-term imaging to track the transplanted cells in recipient SCID mice. Furthermore, we outline a thorough approach to confirm the presence and functional integration of the transplanted cells within the recipient livers.