A novel approach to generate enzyme-free single cell suspensions from archived tissues for miRNA sequencing
Obtaining high-quality omics data at the single-cell level from archived human tissue samples is crucial for gaining insights into cellular heterogeneity and pushing the field of personalized medicine forward. In this technical brief we present a comprehensive methodological framework for the effici...
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| Hauptverfasser: | , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
June 2024
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| In: |
SLAS technology
Year: 2024, Jahrgang: 29, Heft: 3, Pages: 1-7 |
| ISSN: | 2472-6311 |
| DOI: | 10.1016/j.slast.2024.100133 |
| Online-Zugang: | Verlag, kostenfrei, Volltext: https://doi.org/10.1016/j.slast.2024.100133 Verlag, kostenfrei, Volltext: https://www.sciencedirect.com/science/article/pii/S2472630324000153 |
| Verfasserangaben: | Stefan Scheuermann, Sarah Hücker, Annika Engel, Nicole Ludwig, Philipp Lebhardt, Jens Langejürgen, Stefan Kirsch |
| Zusammenfassung: | Obtaining high-quality omics data at the single-cell level from archived human tissue samples is crucial for gaining insights into cellular heterogeneity and pushing the field of personalized medicine forward. In this technical brief we present a comprehensive methodological framework for the efficient enzyme-free preparation of tissue-derived single cell suspensions and their conversion into single-cell miRNA sequencing libraries. The resulting data from this study have the potential to deepen our understanding of miRNA expression at the single-cell level and its relevance in the context of the examined tissues. The workflow encompasses tissue collection, RNALater immersion, storage, thawing, TissueGrinder-mediated dissociation, miRNA lysis, library preparation, sequencing, and data analysis. Quality control measures ensure reliable miRNA data, with specific attention to sample quality. The UMAP analysis reveals tissue-specific cell clustering, while miRNA diversity reflects tissue variations. The presented workflow effectively processes preserved tissues, extending opportunities for retrospective analysis and biobank utilization. |
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| Beschreibung: | Online verfügbar: 6. April 2024, Artikelversion: 8. Juni 2024 Gesehen am 21.01.2025 |
| Beschreibung: | Online Resource |
| ISSN: | 2472-6311 |
| DOI: | 10.1016/j.slast.2024.100133 |