Diffusion Limited Cryopreservation of Tissue with Radiofrequency Heated Metal Forms

Zonghu Han, Anirudh Sharma, Zhe Gao, Timothy W. Carlson, M. Gerard O'Sullivan, Erik B. Finger, John C. Bischof

Research output: Contribution to journalArticlepeer-review

Abstract

Cryopreserved tissues are increasingly needed in biomedical applications. However, successful cryopreservation is generally only reported for thin tissues (≤1 mm). This work presents several innovations to reduce cryoprotectant (CPA) toxicity and improve tissue cryopreservation, including 1) improved tissue warming rates through radiofrequency metal form and field optimization and 2) an experimentally verified predictive model to optimize CPA loading and rewarming to reduce toxicity. CPA loading is studied by microcomputed tomography (µCT) imaging, rewarming by thermal measurements, and modeling, and viability is measured after loading and/or cryopreservation by alamarBlue and histology. Loading conditions for three common CPA cocktails (6, 8.4, and 9.3 m) are designed, and then fast cooling and metal forms rewarming (up to 2000 °C min−1) achieve ≥90% viability in cryopreserved 1–2 mm arteries with various CPAs. Despite high viability by alamarBlue, histology shows subtle changes after cryopreservation suggesting some degree of cell damage especially in the central portions of thicker arteries up to 2 mm. While further studies are needed, these results show careful CPA loading and higher metal forms warming rates can help reduce CPA loading toxicity and improve outcomes from cryopreservation in tissues while also offering new protocols to preserve larger tissues ≥1 mm in thickness.

Original languageEnglish (US)
Article number2000796
JournalAdvanced Healthcare Materials
Volume9
Issue number19
DOIs
StatePublished - Oct 1 2020

Bibliographical note

Funding Information:
A.S. and Z.G. contributed equally to this work. This work was supported by NIH R01HL135046 and R01DK117425‐01. The Visible Heart Laboratory's Alex Deakyne is thanked for access to porcine arteries.

Publisher Copyright:
© 2020 The Authors. Published by Wiley-VCH GmbH

Keywords

  • 2 mm thick tissue systems
  • porcine aortas
  • tissue cryopreservation

PubMed: MeSH publication types

  • Journal Article
  • Research Support, N.I.H., Extramural

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