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Related Experiment Video

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Visualization of Cell Cycle Variations and Determination of Nucleation in Postnatal Cardiomyocytes
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Protocol for quantifying murine cardiomyocyte cell division by single-cell suspension.

Samantha K Swift1, Alexandra L Purdy1, Michaela Patterson2

  • 1Department of Cell Biology, Neurobiology, and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA.

STAR Protocols
|November 9, 2024
PubMed
Summary

This study introduces a simple rodent protocol to confirm cardiomyocyte cell division in vivo. The method uses thymidine analogs and fluorescent imaging to analyze ploidy, enabling researchers to track cardiac regeneration.

Keywords:
Cell BiologyDevelopmental biologyMicroscopy

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Area of Science:

  • Cardiovascular Biology
  • Developmental Biology
  • Regenerative Medicine

Background:

  • Current methods for studying cardiomyocyte division in vivo are limited.
  • Distinguishing true cell division from other outcomes is challenging.
  • A need exists for accessible techniques in cardiac regeneration research.

Purpose of the Study:

  • To present a low-barrier protocol for determining cardiomyocyte cell division in rodents.
  • To enable researchers to reliably assess cardiomyocyte proliferation in vivo.
  • To provide a method for retrospective analysis of cell division.

Main Methods:

  • Administration of thymidine analogs to label dividing cells.
  • Langendorff procedure for tissue preservation.
  • Immunofluorescent labeling and microscopy for ploidy assessment.
  • Analysis of fluorescent images to confirm cell division.

Main Results:

  • The protocol allows for retrospective determination of cardiomyocyte cell division.
  • It enables quantification of total and cycling cardiomyocytes.
  • Cellular dimensions can also be measured.

Conclusions:

  • This protocol offers a straightforward approach to study cardiomyocyte division.
  • It addresses a critical gap in cardiac regeneration and development research.
  • The method facilitates robust assessment of cardiomyocyte proliferation in vivo.