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Isolation and Characterization of Adult Cardiac Fibroblasts and Myofibroblasts
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Fibroblast and Myofibroblast Subtypes: Single Cell Sequencing.

Hesham Soliman1,2, Lin Wei Tung1, Fabio M V Rossi3

  • 1School of Biomedical Engineering and Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada.

Methods in Molecular Biology (Clifton, N.J.)
|May 24, 2021
PubMed
Summary

New single-cell technologies reveal diverse fibroblast subtypes within the heart stroma. This research details methods for analyzing this cellular heterogeneity to understand fibroblast roles in organ health and disease.

Keywords:
10× GenomicsComputational analysisFibroblasts and myofibroblast subsetsHeartSingle cell RNA sequencingSpatial transcriptomicsVisium

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Area of Science:

  • Cardiovascular Biology
  • Cellular Heterogeneity
  • Single-Cell Omics

Background:

  • The organ stroma, primarily composed of fibroblasts and myofibroblasts, is critical for tissue structure and repair.
  • Fibroblast heterogeneity has been historically challenging to study due to limited analytical tools.
  • Advancements in single-cell omics technologies offer unprecedented resolution for characterizing cellular diversity.

Purpose of the Study:

  • To explore the transcriptional, functional, epigenetic, and spatial heterogeneity of cardiac fibroblasts and myofibroblasts.
  • To establish robust methods for single-cell RNA sequencing and spatial transcriptomics of heart tissue.
  • To integrate multi-omics data for comprehensive characterization of fibroblast subtypes and their localization.

Main Methods:

  • Isolation and preparation of single-cell suspensions from heart tissue for single-cell RNA sequencing.
  • Preparation of high-quality tissue sections for spatial transcriptomics.
  • Computational analysis and integration of single-cell RNA sequencing and spatial transcriptomics data.

Main Results:

  • Identification of transcriptionally distinct cell clusters within the cardiac stroma.
  • Characterization of fibroblast and myofibroblast heterogeneity at single-cell resolution.
  • Mapping the spatial distribution of identified fibroblast subpopulations within the heart.

Conclusions:

  • Single-cell omics approaches are revolutionizing the understanding of stromal cell heterogeneity.
  • This study provides a framework for dissecting fibroblast diversity and localization in the heart.
  • Further research is needed to determine if identified clusters represent distinct subpopulations or dynamic states.