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Optimized Methods for scRNA-seq and snRNA-seq of Skeletal Muscle Stored in Nucleic Acid Stabilizing Preservative

GSE282362 Homo sapiens Expression profiling by high throughput sequencing 4 samples Submitted 2024/11/21 Platform GPL34281
Summary
Single cell studies have transformed our understanding of cellular heterogeneity in disease but the need for fresh starting material can be an obstacle, especially in the context of international multicenter studies and archived tissue. We developed a protocol to obtain high-quality cells and nuclei from dissected human skeletal muscle archived in the preservative Allprotect® Tissue Reagent. After fluorescent imaging microscopy confirmed intact nuclei, we performed four protocol variations that compared sequencing metrics between cells and nuclei enriched by either filtering or flow cytometry sorting. Cells and nuclei (either sorted or filtered) produced statistically identical transcriptional profiles and recapitulated 8 cell types present in skeletal muscle. Flow cytometry sorting successfully enriched for higher-quality cells and nuclei but resulted in an overall decrease in input material. Our protocol provides an important resource for obtaining high-quality single cell genomic material from archived tissue and to streamline global collaborative efforts.
Published in
Optimized methods for scRNA-seq and snRNA-seq of skeletal muscle stored in nucleic acid stabilizing preservative
Heuston EF, Doumatey AP, Naz F et al. · Communications biology 2025 · PMID 39755918 · doi:10.1038/s42003-024-07445-2
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Also filed as BioProject PRJNA1188202 and SRA study SRP546272. Searching any of these in the dataset finder brings you back here.

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