Physicochemical Mechanisms of Protection Offered by Agarose Encapsulation during Cryopreservation of Mammalian Cells in the Absence of Membrane-Penetrating Cryoprotectants

Mian Wang, Advitiya Mahajan, Jeffrey S. Miller, David H. McKenna, Alptekin Aksan

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

During freeze/thaw, cells are exposed to mechanical, thermal, chemical, and osmotic stresses, which cause loss of viability and function. Cryopreservation agents such as dimethyl sulfoxide (DMSO) are deployed to minimize freeze/thaw damage. However, there is a pressing need to eliminate DMSO from cryopreservation solutions due to its adverse effects. This is of the highest priority especially for cryopreservation of infusible/transplantable cell therapy products. In order to address this issue, we introduce reversible encapsulation in agarose hydrogels in the presence of the membrane-impermeable cryoprotectant, trehalose, as a viable, safe, and effective cryopreservation method. Our findings, which are supported by IR spectroscopy and differential scanning calorimetry analyses, demonstrate that encapsulation in 0.75% agarose hydrogels containing 10-20% trehalose inhibits mechanical damage induced by eutectic phase change, devitrification, and recrystallization, resulting in post-thaw viability comparable to the gold standard 10% DMSO.

Original languageEnglish (US)
Pages (from-to)2226-2236
Number of pages11
JournalACS Applied Bio Materials
Volume6
Issue number6
DOIs
StatePublished - Jun 19 2023

Bibliographical note

Publisher Copyright:
© 2023 American Chemical Society.

Keywords

  • DMSO
  • cryopreservation
  • devitrification
  • encapsulation
  • eutectic melting
  • recrystallization
  • trehalose

PubMed: MeSH publication types

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

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