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<p>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&#xa0;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.</p>

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

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年2月15日

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  • 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

摘要

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.