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Updated: Jan 12, 2026
Isolation of Regenerating Hepatocytes after Partial Hepatectomy in Mice
Published on: December 2, 2022
Isolation and characterisation of AFP+/DLK1+ double-positive hepatic stem/progenitor-like cells from fibrotic partial
Shuangliang Sun1, Tegshjargal Badamjav1,2, Namur Narid1
1State Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Science, Inner Mongolia University, Hohhot, 010031, Inner Mongolia Autonomous Region, China.
Background:
The liver is an organ with a strong regenerative capacity. Currently, partial hepatectomy (PHx) remains the most effective treatment option within various therapeutic strategies for liver diseases. When the liver undergoes acute injury, such as PHx, the remaining hepatocytes become hypertrophic, re-enter the cell cycle, initiate proliferation, and restore liver function. However, identifying the role of adult hepatic stem/progenitor cells (HSPCs) in this repair process remains challenging.
Methods:
In this study, liver fibrosis was induced in mice by subcutaneous injection of carbon tetrachloride (CCl4) into the back for 8 weeks. Subsequently, 2/3 PHx was used to establish a fibrotic PHx model. Samples were collected at 12 h, 24 h, 48 h, 4 d, and 7 d after PHx. The expression of HSPC-associated markers, including Afp, Dlk1, Ki67, and Lgr5, was analysed using quantitative real-time PCR (qRT-PCR) and immunofluorescence.
Results:
Gene expression analysis revealed that the higher concentration of AFP⁺/DLK1⁺ double-positive cells was in the 50-70% Percoll layer, isolated via density gradient differential centrifugation. Pre-injection of CCl4 promoted liver regeneration after 2/3 PHx, with a peak regenerative phase observed at 24-48 h. Experimental results demonstrated that AFP+/DLK1+ double-positive cells exhibit stem cell-like properties and are involved in liver regeneration. Interestingly, adult and embryonic HSPCs were found in distinct Percoll layers and required different in vitro culture conditions.
Conclusions:
AFP and DLK1 serve as potential markers for selecting HSPC-like populations from the fibrotic PHx model. Our findings indicate that the fibrotic liver exhibits the most vigorous regenerative activity at 48 h after PHx, making it the optimal period for the isolation and extraction of HSPC-like cells which refers only to marker expression and proliferation, the relevance of the findings presented herein remains to be further verified.
