Isolating Nuclei From Frozen Human Heart Tissue for Single-Nucleus RNA Sequencing
Date
2022
Authors
Safabakhsh, S.
Sar, F.
Martelotto, L.
Haegert, A.
Singhera, G.
Hanson, P.
Parker, J.
Collins, C.
Rohani, L.
Laksman, Z.
Editors
Advisors
Journal Title
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Type:
Journal article
Citation
Current Protocols in Neuroscience, 2022; 2(7):e480-e480
Statement of Responsibility
Sina Safabakhsh, Funda Sar, Luciano Martelotto, Anne Haegert, Gurpreet Singhera, Paul Hanson, Jeremy Parker, Colin Collins, Leili Rohani, Zachary Laksman
Conference Name
DOI
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.
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Dissertation Note
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Description
Version of Record online: 11 July 2022
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© 2022 Wiley Periodicals LLC.