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Updated: Jun 19, 2026
Isolation of Primary Mouse Hepatocytes for Nascent Protein Synthesis Analysis by Non-radioactive L-azidohomoalanine Labeling Method
Published on: October 23, 2018
Identification and location of label retaining cells in mouse liver
Fusheng Li1, Leilei Lu, Jingjing Lu
1Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, 310018 Hangzhou, People's Republic of China. lfscn@163.com
Background:
In most somatic tissues, adult stem cells are crucial for the maintenance of tissue homeostasis under normal physiological states and during recovery from injuries. Label retaining cell (LRC) assay remains the well-known method to identify possible somatic stem/progenitor cells and their location both in situ and in vivo.
Methods:
Here, BrdU was used to tag the possible hepatic stem/progenitor cells (HSPCs) in newborn pups, followed by a trace period of up to 23 months. Additionally, we report a method to rapidly kill proliferating cells in adult liver tissue, and activate and label (KAL) surviving possible HSPCs.
Results:
We found that LRCs definitively exist in the liver tissues of adult mice, that LRCs express cell cycle proteins cyclind3 and cdk6, but do not express sca-1 or c-kit, and that LRCs locate primarily in the periportal and pericentral regions. Moreover, the number of these LRCs remains nearly constant during the lifespan of the mice. After injury induced by 5-fluorouracil, we observed that the activation of possible HSPCs tagged by the BrdU label was almost completely inhibited at day 4. The cellular kinetics of repair of BrdU-tagged HSPCs were different every 12 h between day 3 and day 4. Moreover, HSPCs still retained labels and located definitively in the periportal region after a prolonged chase.
Conclusions:
The LRC method together with our novel KAL method reported here may be used to identify and locate possible HSPCs.
