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Methods for isolation and transcriptional profiling of individual cells from the human heart

Neha Pimpalwar, Tomasz Czuba, Maya Landenhed Smith, Johan Nilsson, Olof Gidlöf, J. G. Smith

2020Heliyon21 citationsDOIOpen Access PDF

Abstract

BACKGROUND: Global transcriptional profiling of individual cells represents a powerful approach to systematically survey contributions from cell-specific molecular phenotypes to human disease states but requires tissue-specific protocols. Here we sought to comprehensively evaluate protocols for single cell isolation and transcriptional profiling from heart tissue, focusing particularly on frozen tissue which is necessary for study of human hearts at scale. METHODS AND RESULTS: nuclei) from the same human heart. CONCLUSION: Our results confirm the validity of single-nucleus but not single-cell isolation for transcriptional profiling of individual cells from frozen heart tissue, and establishes PCM1-gating as an efficient tool for cardiomyocyte depletion. In addition, our results provide a perspective of cell types inferred from single-nucleus transcriptomes that are present in an adult human heart.

Topics & Concepts

TranscriptomeFlow cytometryBiologyCell biologyCellRNAMolecular biologyGene expressionGeneBiochemistryCongenital heart defects researchSingle-cell and spatial transcriptomicsPluripotent Stem Cells Research
Methods for isolation and transcriptional profiling of individual cells from the human heart | Litcius