Partial freezing of rat livers extends preservation time by 5-fold

Shannon N. Tessier; Reinier J. de Vries; Casie A. Pendexter; Stephanie E.J. Cronin; Sinan Ozer; Ehab O.A. Hafiz; Siavash Raigani; Joao Paulo Oliveira-Costa; Benjamin T. Wilks; Manuela Lopera Higuita; Thomas M. van Gulik; Osman Berk Usta; Shannon L. Stott; Heidi Yeh; Martin L. Yarmush; Korkut Uygun; Mehmet Toner

doi:https://doi.org/10.1038/s41467-022-31490-2

Partial freezing of rat livers extends preservation time by 5-fold

Shannon N. Tessier, Reinier J. de Vries, Casie A. Pendexter, Stephanie E.J. Cronin, Sinan Ozer, Ehab O.A. Hafiz, Siavash Raigani, Joao Paulo Oliveira-Costa, Benjamin T. Wilks, Manuela Lopera Higuita, Thomas M. van Gulik, Osman Berk Usta, Shannon L. Stott, Heidi Yeh, Martin L. Yarmush, Korkut Uygun, Mehmet Toner

School of Engineering, Biomedical Engineering

Rutgers, The State University

Research output: Contribution to journal › Article › peer-review

Abstract

The limited preservation duration of organs has contributed to the shortage of organs for transplantation. Recently, a tripling of the storage duration was achieved with supercooling, which relies on temperatures between −4 and −6 °C. However, to achieve deeper metabolic stasis, lower temperatures are required. Inspired by freeze-tolerant animals, we entered high-subzero temperatures (−10 to −15 °C) using ice nucleators to control ice and cryoprotective agents (CPAs) to maintain an unfrozen liquid fraction. We present this approach, termed partial freezing, by testing gradual (un)loading and different CPAs, holding temperatures, and storage durations. Results indicate that propylene glycol outperforms glycerol and injury is largely influenced by storage temperatures. Subsequently, we demonstrate that machine perfusion enhancements improve the recovery of livers after freezing. Ultimately, livers that were partially frozen for 5-fold longer showed favorable outcomes as compared to viable controls, although frozen livers had lower cumulative bile and higher liver enzymes.

Original language	English (US)
Article number	4008
Journal	Nature communications
Volume	13
Issue number	1
DOIs	https://doi.org/10.1038/s41467-022-31490-2
State	Published - Dec 2022

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
General
Physics and Astronomy(all)

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https://doi.org/10.1038/s41467-022-31490-2

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Tessier, S. N., de Vries, R. J., Pendexter, C. A., Cronin, S. E. J., Ozer, S., Hafiz, E. O. A., Raigani, S., Oliveira-Costa, J. P., Wilks, B. T., Lopera Higuita, M., van Gulik, T. M., Usta, O. B., Stott, S. L., Yeh, H., Yarmush, M. L., Uygun, K., & Toner, M. (2022). Partial freezing of rat livers extends preservation time by 5-fold. Nature communications, 13(1), [4008]. https://doi.org/10.1038/s41467-022-31490-2

@article{c115e97da0be4453a8e44ab361e9b9ec,

title = "Partial freezing of rat livers extends preservation time by 5-fold",

abstract = "The limited preservation duration of organs has contributed to the shortage of organs for transplantation. Recently, a tripling of the storage duration was achieved with supercooling, which relies on temperatures between −4 and −6 °C. However, to achieve deeper metabolic stasis, lower temperatures are required. Inspired by freeze-tolerant animals, we entered high-subzero temperatures (−10 to −15 °C) using ice nucleators to control ice and cryoprotective agents (CPAs) to maintain an unfrozen liquid fraction. We present this approach, termed partial freezing, by testing gradual (un)loading and different CPAs, holding temperatures, and storage durations. Results indicate that propylene glycol outperforms glycerol and injury is largely influenced by storage temperatures. Subsequently, we demonstrate that machine perfusion enhancements improve the recovery of livers after freezing. Ultimately, livers that were partially frozen for 5-fold longer showed favorable outcomes as compared to viable controls, although frozen livers had lower cumulative bile and higher liver enzymes.",

author = "Tessier, {Shannon N.} and {de Vries}, {Reinier J.} and Pendexter, {Casie A.} and Cronin, {Stephanie E.J.} and Sinan Ozer and Hafiz, {Ehab O.A.} and Siavash Raigani and Oliveira-Costa, {Joao Paulo} and Wilks, {Benjamin T.} and {Lopera Higuita}, Manuela and {van Gulik}, {Thomas M.} and Usta, {Osman Berk} and Stott, {Shannon L.} and Heidi Yeh and Yarmush, {Martin L.} and Korkut Uygun and Mehmet Toner",

note = "Funding Information: This research was funded from the US National Institutes of Health, including R01DK114506 (M.T., K.U., M.L.Y.), R01DK096075 (K.U., H.Y.), R01DK107875 (K.U.), R01EB028782 (KU), and National Science Foundation under Grant No. EEC 1941543 (M.T.). Further, we gratefully acknowledge funding to SNT from NIH (K99/R00 HL1431149; R01HL157803), American Heart Association (18CDA34110049), Harvard Medical School Eleanor and Miles Shore Fellowship, and the Claflin Distinguished Scholar Award on behalf of the MGH Executive Committee on Research. We thankfully acknowledge support provided by the Tosteson Fellowship awarded to R.J.V. by the Executive Committee on Research at the Massachusetts General Hospital. We thank the MGH Cell Resource Core, including Peony D. Banik and Sonal Nagpal, for assistance with perfusion equipment and the Mass Spectrometry Special Shared Facilities at Shriners Hospitals for Children (Boston) for quantification of tissue adenylate levels. Finally, we are grateful for our collaboration on high subzero organ preservation with Sylvatica Biotech, Inc and acknowledge awards R44 DK124053-01 (US NIH), W81XWH21C0060 (US Dept. of Defense) and support from ATCC. Funding Information: This research was funded from the US National Institutes of Health, including R01DK114506 (M.T., K.U., M.L.Y.), R01DK096075 (K.U., H.Y.), R01DK107875 (K.U.), R01EB028782 (KU), and National Science Foundation under Grant No. EEC 1941543 (M.T.). Further, we gratefully acknowledge funding to SNT from NIH (K99/R00 HL1431149; R01HL157803), American Heart Association (18CDA34110049), Harvard Medical School Eleanor and Miles Shore Fellowship, and the Claflin Distinguished Scholar Award on behalf of the MGH Executive Committee on Research. We thankfully acknowledge support provided by the Tosteson Fellowship awarded to R.J.V. by the Executive Committee on Research at the Massachusetts General Hospital. We thank the MGH Cell Resource Core, including Peony D. Banik and Sonal Nagpal, for assistance with perfusion equipment and the Mass Spectrometry Special Shared Facilities at Shriners Hospitals for Children (Boston) for quantification of tissue adenylate levels. Finally, we are grateful for our collaboration on high subzero organ preservation with Sylvatica Biotech, Inc and acknowledge awards R44 DK124053-01 (US NIH), W81XWH21C0060 (US Dept. of Defense) and support from ATCC. Publisher Copyright: {\textcopyright} 2022, The Author(s).",

year = "2022",

month = dec,

doi = "https://doi.org/10.1038/s41467-022-31490-2",

language = "English (US)",

volume = "13",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

number = "1",

}

Tessier, SN, de Vries, RJ, Pendexter, CA, Cronin, SEJ, Ozer, S, Hafiz, EOA, Raigani, S, Oliveira-Costa, JP, Wilks, BT, Lopera Higuita, M, van Gulik, TM, Usta, OB, Stott, SL, Yeh, H, Yarmush, ML, Uygun, K & Toner, M 2022, 'Partial freezing of rat livers extends preservation time by 5-fold', Nature communications, vol. 13, no. 1, 4008. https://doi.org/10.1038/s41467-022-31490-2

TY - JOUR

T1 - Partial freezing of rat livers extends preservation time by 5-fold

AU - Tessier, Shannon N.

AU - de Vries, Reinier J.

AU - Pendexter, Casie A.

AU - Cronin, Stephanie E.J.

AU - Ozer, Sinan

AU - Hafiz, Ehab O.A.

AU - Raigani, Siavash

AU - Oliveira-Costa, Joao Paulo

AU - Wilks, Benjamin T.

AU - Lopera Higuita, Manuela

AU - van Gulik, Thomas M.

AU - Usta, Osman Berk

AU - Stott, Shannon L.

AU - Yeh, Heidi

AU - Yarmush, Martin L.

AU - Uygun, Korkut

AU - Toner, Mehmet

N1 - Funding Information: This research was funded from the US National Institutes of Health, including R01DK114506 (M.T., K.U., M.L.Y.), R01DK096075 (K.U., H.Y.), R01DK107875 (K.U.), R01EB028782 (KU), and National Science Foundation under Grant No. EEC 1941543 (M.T.). Further, we gratefully acknowledge funding to SNT from NIH (K99/R00 HL1431149; R01HL157803), American Heart Association (18CDA34110049), Harvard Medical School Eleanor and Miles Shore Fellowship, and the Claflin Distinguished Scholar Award on behalf of the MGH Executive Committee on Research. We thankfully acknowledge support provided by the Tosteson Fellowship awarded to R.J.V. by the Executive Committee on Research at the Massachusetts General Hospital. We thank the MGH Cell Resource Core, including Peony D. Banik and Sonal Nagpal, for assistance with perfusion equipment and the Mass Spectrometry Special Shared Facilities at Shriners Hospitals for Children (Boston) for quantification of tissue adenylate levels. Finally, we are grateful for our collaboration on high subzero organ preservation with Sylvatica Biotech, Inc and acknowledge awards R44 DK124053-01 (US NIH), W81XWH21C0060 (US Dept. of Defense) and support from ATCC. Funding Information: This research was funded from the US National Institutes of Health, including R01DK114506 (M.T., K.U., M.L.Y.), R01DK096075 (K.U., H.Y.), R01DK107875 (K.U.), R01EB028782 (KU), and National Science Foundation under Grant No. EEC 1941543 (M.T.). Further, we gratefully acknowledge funding to SNT from NIH (K99/R00 HL1431149; R01HL157803), American Heart Association (18CDA34110049), Harvard Medical School Eleanor and Miles Shore Fellowship, and the Claflin Distinguished Scholar Award on behalf of the MGH Executive Committee on Research. We thankfully acknowledge support provided by the Tosteson Fellowship awarded to R.J.V. by the Executive Committee on Research at the Massachusetts General Hospital. We thank the MGH Cell Resource Core, including Peony D. Banik and Sonal Nagpal, for assistance with perfusion equipment and the Mass Spectrometry Special Shared Facilities at Shriners Hospitals for Children (Boston) for quantification of tissue adenylate levels. Finally, we are grateful for our collaboration on high subzero organ preservation with Sylvatica Biotech, Inc and acknowledge awards R44 DK124053-01 (US NIH), W81XWH21C0060 (US Dept. of Defense) and support from ATCC. Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - The limited preservation duration of organs has contributed to the shortage of organs for transplantation. Recently, a tripling of the storage duration was achieved with supercooling, which relies on temperatures between −4 and −6 °C. However, to achieve deeper metabolic stasis, lower temperatures are required. Inspired by freeze-tolerant animals, we entered high-subzero temperatures (−10 to −15 °C) using ice nucleators to control ice and cryoprotective agents (CPAs) to maintain an unfrozen liquid fraction. We present this approach, termed partial freezing, by testing gradual (un)loading and different CPAs, holding temperatures, and storage durations. Results indicate that propylene glycol outperforms glycerol and injury is largely influenced by storage temperatures. Subsequently, we demonstrate that machine perfusion enhancements improve the recovery of livers after freezing. Ultimately, livers that were partially frozen for 5-fold longer showed favorable outcomes as compared to viable controls, although frozen livers had lower cumulative bile and higher liver enzymes.

AB - The limited preservation duration of organs has contributed to the shortage of organs for transplantation. Recently, a tripling of the storage duration was achieved with supercooling, which relies on temperatures between −4 and −6 °C. However, to achieve deeper metabolic stasis, lower temperatures are required. Inspired by freeze-tolerant animals, we entered high-subzero temperatures (−10 to −15 °C) using ice nucleators to control ice and cryoprotective agents (CPAs) to maintain an unfrozen liquid fraction. We present this approach, termed partial freezing, by testing gradual (un)loading and different CPAs, holding temperatures, and storage durations. Results indicate that propylene glycol outperforms glycerol and injury is largely influenced by storage temperatures. Subsequently, we demonstrate that machine perfusion enhancements improve the recovery of livers after freezing. Ultimately, livers that were partially frozen for 5-fold longer showed favorable outcomes as compared to viable controls, although frozen livers had lower cumulative bile and higher liver enzymes.

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DO - https://doi.org/10.1038/s41467-022-31490-2

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VL - 13

JO - Nature communications

JF - Nature communications

SN - 2041-1723

IS - 1

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ER -

Partial freezing of rat livers extends preservation time by 5-fold

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