Please use this identifier to cite or link to this item:
Scopus Web of Science® Altmetric
Type: Journal article
Title: Isolating Nuclei From Frozen Human Heart Tissue for Single-Nucleus RNA Sequencing
Author: Safabakhsh, S.
Sar, F.
Martelotto, L.
Haegert, A.
Singhera, G.
Hanson, P.
Parker, J.
Collins, C.
Rohani, L.
Laksman, Z.
Citation: Current Protocols, 2022; 2(7):e480-e480
Publisher: Wiley
Issue Date: 2022
ISSN: 2691-1299
Statement of
Sina Safabakhsh, Funda Sar, Luciano Martelotto, Anne Haegert, Gurpreet Singhera, Paul Hanson, Jeremy Parker, Colin Collins, Leili Rohani, Zachary Laksman
Abstract: Heart disease is the leading cause of global morbidity and mortality. This is in part because, despite an abundance of animal and in vitro models, it has been a challenge to date to study human heart tissue with sufficient depth and resolution to develop disease-modifying therapies for common cardiac conditions. Single-nucleus RNA sequencing (snRNA-seq) has emerged as a powerful tool capable of analyzing cellular function and signaling in health and disease, and has already contributed to significant advances in areas such as oncology and hematology. Employing snRNA-seq technology on flash-frozen human tissue has the potential to unlock novel disease mechanisms and pathways in any organ. Studying the human heart using snRNA-seq is a key priority for the field of cardiovascular sciences; however, progress to date has been slowed by numerous barriers. One key challenge is the fact that the human heart is very resistant to shearing and stress, making tissue dissociation and nuclear isolation difficult. Here, we describe a tissue dissociation method allowing the efficient and cost-effective isolation of high-quality nuclei from flash-frozen human heart tissue collected in surgical operating rooms. Our protocol addresses the challenge of nuclear isolation from human hearts, enables snRNA-seq of the human heart, and paves the way for an improved understanding of the human heart in health and disease. Ultimately, this will be key to uncovering signaling pathways and networks amenable to therapeutic intervention and the development of novel biomarkers and disease-modifying therapies.
Keywords: cardiac
Description: Version of Record online: 11 July 2022
Rights: © 2022 Wiley Periodicals LLC.
DOI: 10.1002/cpz1.480
Appears in Collections:Medicine publications
South Australian Immunogenomics Cancer Institute (SAIGENCI) publications

Files in This Item:
There are no files associated with this item.

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.