Optimized methods for scRNA-seq and snRNA-seq of skeletal muscle stored in nucleic acid stabilizing preservative
2025

Optimized methods for single cell RNA sequencing of preserved skeletal muscle

Sample size: 8 publication 10 minutes Evidence: high

Author Information

Author(s): Heuston Elisabeth F., Doumatey Ayo P., Naz Faiza, Islam Shamima, Anderson Stacie, Kirby Martha R., Wincovitch Stephen, Dell’Orso Stefania, Rotimi Charles N., Adeyemo Adebowale A.

Primary Institution: National Institutes of Health

Hypothesis

Is tissue stored in Allprotect® suitable for single-cell transcription profiling?

Conclusion

The study demonstrates that archived skeletal muscle stored in Allprotect® Tissue Reagent produces high-quality data suitable for single-cell RNA sequencing.

Supporting Evidence

  • All four processing methods yielded similar transcriptional profiles.
  • Flow cytometry sorting enriched for higher-quality cells but reduced input material.
  • Archived muscle tissue in Allprotect® retains the variety of cell types present in vivo.

Takeaway

Researchers found a way to study muscle cells from preserved samples, which helps scientists learn more about diseases without needing fresh tissue.

Methodology

The study developed and compared four processing pipelines for single-cell RNA sequencing using preserved skeletal muscle tissue.

Potential Biases

Potential biases in sample quality due to the preservation method and sorting processes.

Limitations

The method is optimized specifically for human skeletal muscle and may require adjustments for other tissues.

Participant Demographics

Adult Nigerians enrolled in the Africa America Diabetes Mellitus study.

Statistical Information

P-Value

p<0.05

Statistical Significance

p<0.05

Digital Object Identifier (DOI)

10.1038/s42003-024-07445-2

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