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In Vivo Clonal Analysis of Cardiomyocytes

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Cardiac Regeneration

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2158))

Abstract

The development of the CreER/LoxP system has enabled temporal control and cell type specificity of gene activation or repression. A common application of this system involves lineage tracing and examining the proliferative capacity of cells of interest through clonal analysis. Here, we describe a method of performing 2- and 3-dimensional clonal analysis of cardiomyocytes (CMs) using the Rainbow reporter mouse model. We outline the process of using the Cell Counter plug-in tool in ImageJ to quantify the number of clones as well as the number of cells within each clone. For 3-dimensional analysis, we describe the tissue clearing technique, CLARITY, in conjunction with light-sheet imaging to obtain digital slices of the whole heart that can be reconstructed and clone volumes quantified using the Imaris software.

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Acknowledgments

This work was supported by the F31 HL144057 (N.B.N.), NIH DP2 HL127728 and UCLA Broad Stem Cell Research Center-Rose Hills Foundation Research Award (R.A.).

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Authors and Affiliations

  1. Division of Cardiology, Department of Internal Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, USA

    Ngoc B. Nguyen, Yichen Ding, Tzung Hsiai & Reza Ardehali

  2. Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, USA

    Ngoc B. Nguyen & Reza Ardehali

  3. Molecular, Cellular and Integrative Physiology Graduate Program, University of California, Los Angeles, Los Angeles, CA, USA

    Ngoc B. Nguyen & Reza Ardehali

  4. Children’s Hospital Los Angeles and the Saban Research Institute, Los Angeles, CA, USA

    G. Esteban Fernandez

  5. Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA

    Reza Ardehali

Authors
  1. Ngoc B. Nguyen
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  2. G. Esteban Fernandez
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  3. Yichen Ding
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  4. Tzung Hsiai
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  5. Reza Ardehali
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Corresponding author

Correspondence to Reza Ardehali .

Editor information

Editors and Affiliations

  1. Department of Cell Biology, Duke University Medical Center, Durham, NC, USA

    Kenneth D. Poss

  2. Division of Cardiology, UPMC Children’s Hospital of Pittsburgh, Pittsburgh, PA, USA

    Bernhard Kühn

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Nguyen, N.B., Fernandez, G.E., Ding, Y., Hsiai, T., Ardehali, R. (2021). In Vivo Clonal Analysis of Cardiomyocytes. In: Poss, K.D., Kühn, B. (eds) Cardiac Regeneration. Methods in Molecular Biology, vol 2158. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0668-1_18

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  • DOI: https://doi.org/10.1007/978-1-0716-0668-1_18

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0667-4

  • Online ISBN: 978-1-0716-0668-1

  • eBook Packages: Springer Protocols

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