Cryopreservation of Whole Rat Livers by Vitrification and Nanowarming

Anirudh Sharma, Charles Y. Lee, Bat Erdene Namsrai, Zonghu Han, Diane K Tobolt, Joseph Sushil Rao, Zhe Gao, Michael L. Etheridge, Michael Garwood, Mark G. Clemens, John C. Bischof, Erik B. Finger

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Liver cryopreservation has the potential to enable indefinite organ banking. This study investigated vitrification—the ice-free cryopreservation of livers in a glass-like state—as a promising alternative to conventional cryopreservation, which uniformly fails due to damage from ice formation or cracking. Our unique “nanowarming” technology, which involves perfusing biospecimens with cryoprotective agents (CPAs) and silica-coated iron oxide nanoparticles (sIONPs) and then, after vitrification, exciting the nanoparticles via radiofrequency waves, enables rewarming of vitrified specimens fast enough to avoid ice formation and uniformly enough to prevent cracking from thermal stresses, thereby addressing the two main failures of conventional cryopreservation. This study demonstrates the ability to load rat livers with both CPA and sIONPs by vascular perfusion, cool them rapidly to an ice-free vitrified state, and rapidly and homogenously rewarm them. While there was some elevation of liver enzymes (Alanine Aminotransferase) and impaired indocyanine green (ICG) excretion, the nanowarmed livers were viable, maintained normal tissue architecture, had preserved vascular endothelium, and demonstrated hepatocyte and organ-level function, including production of bile and hepatocyte uptake of ICG during normothermic reperfusion. These findings suggest that cryopreservation of whole livers via vitrification and nanowarming has the potential to achieve organ banking for transplant and other biomedical applications.

Original languageEnglish (US)
Pages (from-to)566-577
Number of pages12
JournalAnnals of Biomedical Engineering
Issue number3
StatePublished - Mar 2023

Bibliographical note

Funding Information:
This work was funded by NIH (HL135046, DK117425, DK131209, DK119043, and DK126551) and NSF EEC 1941543. JSR acknowledges the support of the Schulze Diabetes Institute. We acknowledge the assistance of Dr. Mark Sanders of the University of Minnesota Imaging Center for assistance in freeze-substitution experiments.

Publisher Copyright:
© 2022, The Author(s) under exclusive licence to Biomedical Engineering Society.


  • Critical cooling rate
  • Critical warming rate
  • Cryoprotective agent
  • Liver transplant
  • Organ preservation

PubMed: MeSH publication types

  • Journal Article


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