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

ABSTRACT: Single-cell RNA sequencing (scRNA-seq) remains state-of-the-art for transcriptomic cell-mapping. Here, we provide a protocol to generate high-resolution scRNA-seq of rare cardiomyocyte populations (e.g., regenerating/dividing, etc.) from mouse and zebrafish hearts as well as induced pluripotent stem cells, collected in time to achieve detailed transcriptomic insight. We describe the serial steps of viability staining, methanol fixation, storage, and cell sorting to preserve RNA integrity suited for scRNA-seq as well as the quality assessment of the data as shown by examples. For complete details on the use and execution of this protocol, please refer to Bak et al.¹.

SUBMITTER: Ellman DG

PROVIDER: S-EPMC11345562 | biostudies-literature | 2024 Aug

REPOSITORIES: biostudies-literature

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

Ellman Ditte Gry DG Bjerre Frederik Adam FA Bak Sara Thornby ST Mathiesen Sabrina Bech SB Harvald Eva Bang EB Jensen Charlotte Harken CH Andersen Ditte Caroline DC

STAR protocols 20240801 3

Single-cell RNA sequencing (scRNA-seq) remains state-of-the-art for transcriptomic cell-mapping. Here, we provide a protocol to generate high-resolution scRNA-seq of rare cardiomyocyte populations (e.g., regenerating/dividing, etc.) from mouse and zebrafish hearts as well as induced pluripotent stem cells, collected in time to achieve detailed transcriptomic insight. We describe the serial steps of viability staining, methanol fixation, storage, and cell sorting to preserve RNA integrity suited ...[more]

PMID: 39096494

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