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Application of EdU-Based DNA Synthesis Assay to Measure Hepatocyte Proliferation In Situ During Liver Regeneration

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Hepatocytes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2544))

Abstract

Monitoring hepatocyte proliferation in situ following partial hepatectomy is widely used to characterize cytokines, growth factors and signaling molecules and pathways as well as the regulatory mechanisms involved in liver regeneration. Periodic measurement of the liver/body mass ratio estimates the rate of liver regeneration, which is often supplemented by evaluating the proportion of proliferating hepatocytes using a synthetic nucleoside analog such as 5-bromo-2'-deoxyuridine (BrdU) or the nuclear accumulation of proliferating cell nuclear antigen (PCNA) in proliferating cells. The introduction of the thymidine analog 5-ethynyl-2’deoxyuridine (EdU) and its detection by “click chemistry” using fluorescently labeled reagents has simplified the evaluation of live cell proliferation as it eliminates certain limitations of antibody-mediated detection of BrdU. Here, we describe the EdU-based measurement of hepatocyte proliferation during liver regeneration and correlate the results with that of Ki67 and PCNA-based assays.

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Acknowledgment

This work was supported by the Canadian Institutes of Health Research (CIHR) Project grant to SI (PJT-153174). AG and MN are recipients of postdoctoral fellowship from the Le Fonds de recherche du Québec – Santé (FRQS).

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Authors and Affiliations

  1. Department of Immunology and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, QC, Canada

    Amit Ghosh, Awais Ullah Ihsan, Madhuparna Nandi, Maryse Cloutier, Md Gulam Musawwir Khan, Sheela Ramanathan & Subburaj Ilangumaran

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  1. Amit Ghosh
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  2. Awais Ullah Ihsan
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  3. Madhuparna Nandi
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  4. Maryse Cloutier
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  5. Md Gulam Musawwir Khan
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  6. Sheela Ramanathan
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  7. Subburaj Ilangumaran
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Corresponding author

Correspondence to Subburaj Ilangumaran .

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Editors and Affiliations

  1. Research Institute for Frontier Medicine, Sapporo Medical University, School of Medicine, Sapporo, Hokkaido, Japan

    Naoki Tanimizu

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ghosh, A. et al. (2022). Application of EdU-Based DNA Synthesis Assay to Measure Hepatocyte Proliferation In Situ During Liver Regeneration. In: Tanimizu, N. (eds) Hepatocytes. Methods in Molecular Biology, vol 2544. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2557-6_14

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  • DOI: https://doi.org/10.1007/978-1-0716-2557-6_14

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2556-9

  • Online ISBN: 978-1-0716-2557-6

  • eBook Packages: Springer Protocols

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