<p>Single-cell and spatial transcriptomic technologies have revolutionized human cell and tissue research. Success of these methods critically depends on the preservation of RNA integrity, cellular viability, and structure of the explanted tissues—features that are impacted by tissue storage conditions, and pose challenges in research settings, where tissues are often collected in the operating room and transported to distant laboratories. In this study, we evaluated the effectiveness of a clinically approved organ transplant solution (TS) for preserving skeletal muscle under refrigerated conditions. Using murine and human skeletal muscle tissues we found that compared to buffered saline, TS consistently maintained higher RNA quality, preserved tissue morphology, and supported successful primary cell culture and spatial transcriptomic analysis of murine and human muscle samples obtained from different surgical collection sites. These findings establish a foundation for the use of a standardized protocol for live muscle tissue preservation for reliable downstream transcriptomic analyses for multicenter research efforts.</p>

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Use of organ transplant solution to preserve skeletal muscle for cellular and spatial transcriptomic analyses

  • Prech Uapinyoying,
  • Young Jae Moon,
  • Aiping Zhang,
  • Pamela Barraza-Flores,
  • Won Lee,
  • Angela N. Viaene,
  • Sanda Alexandrescu,
  • Nhu Chau,
  • Casie A. Genetti,
  • Yi-Wen Chen,
  • Alan H. Beggs,
  • Carsten G. Bönnemann,
  • Monkol Lek,
  • Jyoti K. Jaiswal,
  • Rachel Alvarez,
  • Surajit Bhattacharya,
  • Benjamin D. Cosgrove,
  • Jordi Diaz-Manera,
  • Gustavo Dziewczapolski,
  • Emanuela Gussoni,
  • Andrew Jia,
  • Nikki M. McCormack,
  • Douglas Milay,
  • Steven A. Moore,
  • Jessica L. Orton,
  • Casey Swoboda

摘要

Single-cell and spatial transcriptomic technologies have revolutionized human cell and tissue research. Success of these methods critically depends on the preservation of RNA integrity, cellular viability, and structure of the explanted tissues—features that are impacted by tissue storage conditions, and pose challenges in research settings, where tissues are often collected in the operating room and transported to distant laboratories. In this study, we evaluated the effectiveness of a clinically approved organ transplant solution (TS) for preserving skeletal muscle under refrigerated conditions. Using murine and human skeletal muscle tissues we found that compared to buffered saline, TS consistently maintained higher RNA quality, preserved tissue morphology, and supported successful primary cell culture and spatial transcriptomic analysis of murine and human muscle samples obtained from different surgical collection sites. These findings establish a foundation for the use of a standardized protocol for live muscle tissue preservation for reliable downstream transcriptomic analyses for multicenter research efforts.