A versatile and efficient method to isolate nuclei from low-input cryopreserved tissues for single-nuclei transcriptomics

A versatile and efficient method to isolate nuclei from low-input cryopreserved tissues for single-nuclei transcriptomics

Abstract Clinical samples are vital for understanding diseases, but their scarcity requires refined research methods. Emerging single-cell technologies offer detailed views of tissue heterogeneity but need sufficient fully characterized tissues. We developed an optimized single-nuclei RNA sequencing...

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Main Authors: Cristopher Segovia, Vincent Desrosiers, Fatemeh Khadangi, Karine Robitaille, Victoria Saavedra Armero, Myreille D’Astous, Gabriel Khelifi, Alain Bergeron, Samer Hussein, Maxime Richer, Yohan Bossé, Yves Fradet, Vincent Fradet, Steve Bilodeau
Format: Article
Language:English
Published: Nature Portfolio 2025-02-01
Series:Scientific Reports
Subjects:
Nuclei isolation
Tissue heterogeneity
Transcriptomics
Cancer
Tissue homogenization
Nuclei sorting
Online Access:https://doi.org/10.1038/s41598-025-90070-8
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Summary:Abstract Clinical samples are vital for understanding diseases, but their scarcity requires refined research methods. Emerging single-cell technologies offer detailed views of tissue heterogeneity but need sufficient fully characterized tissues. We developed an optimized single-nuclei RNA sequencing (snRNA-seq) protocol to extract nuclei from just 15 mg of cryopreserved human tissue. Applied to four cancer tissues (brain, bladder, lung, prostate), it profiled 1550–7468 nuclei per tissue, revealing heterogeneity comparable to public single-cell atlases. This method enhances the use and sharing of rare, cryopreserved biospecimens, supporting research where sample quantity is limited and full tissue characterization is needed.
ISSN:2045-2322

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