Protocol to achieve high-resolution single-cell transcriptomics of cardiomyocytes in multiple species

Area of Science:

• Cardiovascular Biology
• Molecular Biology
• Genomics

Background:

• Single-cell RNA sequencing (scRNA-seq) is crucial for understanding cellular heterogeneity.
• Identifying and analyzing rare cell populations, like regenerating cardiomyocytes, presents technical challenges.

Purpose of the Study:

• To establish a robust protocol for high-resolution scRNA-seq of rare cardiomyocyte populations.
• To enable detailed transcriptomic analysis of cardiac cells from mouse, zebrafish, and iPSCs.

Main Methods:

• The protocol involves viability staining, methanol fixation, and cell sorting to preserve RNA integrity.
• Quality assessment of generated scRNA-seq data is demonstrated with examples.

Main Results:

• The protocol successfully generates high-resolution scRNA-seq data from rare cardiomyocyte populations.
• Preserved RNA integrity allows for detailed transcriptomic profiling.

Conclusions:

• This optimized scRNA-seq protocol facilitates the study of rare cardiac cell types.
• The method is applicable to diverse biological systems, including developmental and regenerative contexts.