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Identification and location of label retaining cells in mouse liver

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

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.

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Abbreviations

ASCs:

Adult stem cells

LRCs:

Label retaining cells

HSCs/HPCs:

Hepatic stem/progenitor cells

NPC:

Non-parenchymal cell (s)

5-Fu:

5-Fluorouracil

Ck:

Cytokeratin

Sca-1:

Stem cell antigen-1

BrdU:

5-Bromo-2′-deoxyuridine

DAPI:

4,6-Diamidino-2-phenylindole

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Acknowledgments

Sincere thanks go to Xuedong Wang (Institute of Biochemistry, Zhejiang Sci-Tech University) for help in the use of the Confocal Laser Scanning Microscope. This work was greatly supported by the National Support Program for Science and Technology (No. 2006BAI01B04), by the National Natural Science Foundation (No. 30740015), and by a grant for key projects from the Zhejiang Province Natural Science Foundation (No. Y205449).

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Authors and Affiliations

  1. Institute of Biochemistry, College of Life Sciences, Zhejiang Sci-Tech University, 310018, Hangzhou, People’s Republic of China

    Fusheng Li

  2. Department of Bimolecular Engineering, Tohoku University, Aoba-yama 6-6-11-608, Aoba-ku, Sendai, Miyagi, 980-8579, Japan

    Leilei Lu

  3. Changzhou Tenraid Biotech Company, Room 303, Building A, No. 25 Mid Changjiang Rd, 213022, Changzhou, People’s Republic of China

    Fusheng Li, Leilei Lu & Jingjing Lu

Authors
  1. Fusheng Li
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  2. Leilei Lu
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Correspondence to Fusheng Li.

Additional information

F. Li and L. Lu contributed equally to this work.

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Li, F., Lu, L. & Lu, J. Identification and location of label retaining cells in mouse liver. J Gastroenterol 45, 113–121 (2010). https://doi.org/10.1007/s00535-009-0139-2

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  • DOI: https://doi.org/10.1007/s00535-009-0139-2

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