Natural deep eutectic systems for nature-inspired cryopreservation of cells

Kathlyn Hornberger; Rui Li; Ana Rita C. Duarte; Allison Hubel

doi:10.1002/aic.17085

Natural deep eutectic systems for nature-inspired cryopreservation of cells

Kathlyn Hornberger, Rui Li, Ana Rita C. Duarte, Allison Hubel

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

22 Scopus citations

Abstract

Natural deep eutectic systems (NADES) are emerging as potential cryoprotective agents (CPA) for cell preservation. In this investigation, we develop an optimized CPA formulation using trehalose-glycerol NADES (T:G) diluted in Normosol-R and supplemented with isoleucine. Differential scanning calorimetry (DSC) is used to define the thermophysical properties of NADES-based solutions, and Raman spectroscopy is used to characterize the effect of NADES on ice formation and hydrogen bonding. Jurkat cells are cryopreserved in each solution, and post-thaw cell recovery, apoptosis, and growth are quantified. Raman spectra and heat maps show that NADES suppresses both ice formation and dehydration of the nonfrozen region. Supplementing NADES with isoleucine does not affect the solution's thermophysical properties but significantly improves the cells' survival and proliferation post-thaw. The study indicates that thermophysical properties of CPA solutions alone cannot predict optimal cell survival, suggesting that stabilization of biological structures by CPAs may play a role in successful cryopreservation.

Original language	English (US)
Article number	e17085
Journal	AIChE Journal
Volume	67
Issue number	2
DOIs	https://doi.org/10.1002/aic.17085
State	Published - Feb 2021
Externally published	Yes

Bibliographical note

Funding Information:
European Research Council, Grant/Award Number: ERC‐2016‐CoG 725034; National Institutes of Health, Grant/Award Numbers: R01EB023880, R25HL128372; National Science Foundation, Grant/Award Number: CON‐75851; Project 00074041 Funding information

Funding Information:
This work was supported by the National Institutes of Health (R01EB023880, R25HL128372), National Science Foundation Graduate Research Fellowship (CON-75851, Project 00074041) and ERC-2016-CoG 725034 (ERC Consolidator Grant Des.solve). The confocal light microscopy in this work was carried out using a Zeiss Scope with the assistance of Guillermo Marques at the University Imaging Centers, University of Minnesota. The flow cytometry in this work was carried out at the University of Minnesota Flow Cytometry Resource. The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program.

Funding Information:
This work was supported by the National Institutes of Health (R01EB023880, R25HL128372), National Science Foundation Graduate Research Fellowship (CON‐75851, Project 00074041) and ERC‐2016‐CoG 725034 (ERC Consolidator Grant Des.solve). The confocal light microscopy in this work was carried out using a Zeiss Scope with the assistance of Guillermo Marques at the University Imaging Centers, University of Minnesota. The flow cytometry in this work was carried out at the University of Minnesota Flow Cytometry Resource. The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program.

Publisher Copyright:
© 2020 American Institute of Chemical Engineers

Keywords

Raman spectroscopy
amino acids
cryopreservation
differential scanning calorimetry
natural deep eutectic solvents

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title = "Natural deep eutectic systems for nature-inspired cryopreservation of cells",

abstract = "Natural deep eutectic systems (NADES) are emerging as potential cryoprotective agents (CPA) for cell preservation. In this investigation, we develop an optimized CPA formulation using trehalose-glycerol NADES (T:G) diluted in Normosol-R and supplemented with isoleucine. Differential scanning calorimetry (DSC) is used to define the thermophysical properties of NADES-based solutions, and Raman spectroscopy is used to characterize the effect of NADES on ice formation and hydrogen bonding. Jurkat cells are cryopreserved in each solution, and post-thaw cell recovery, apoptosis, and growth are quantified. Raman spectra and heat maps show that NADES suppresses both ice formation and dehydration of the nonfrozen region. Supplementing NADES with isoleucine does not affect the solution's thermophysical properties but significantly improves the cells' survival and proliferation post-thaw. The study indicates that thermophysical properties of CPA solutions alone cannot predict optimal cell survival, suggesting that stabilization of biological structures by CPAs may play a role in successful cryopreservation.",

keywords = "Raman spectroscopy, amino acids, cryopreservation, differential scanning calorimetry, natural deep eutectic solvents",

author = "Kathlyn Hornberger and Rui Li and Duarte, {Ana Rita C.} and Allison Hubel",

note = "Funding Information: European Research Council, Grant/Award Number: ERC‐2016‐CoG 725034; National Institutes of Health, Grant/Award Numbers: R01EB023880, R25HL128372; National Science Foundation, Grant/Award Number: CON‐75851; Project 00074041 Funding information Funding Information: This work was supported by the National Institutes of Health (R01EB023880, R25HL128372), National Science Foundation Graduate Research Fellowship (CON-75851, Project 00074041) and ERC-2016-CoG 725034 (ERC Consolidator Grant Des.solve). The confocal light microscopy in this work was carried out using a Zeiss Scope with the assistance of Guillermo Marques at the University Imaging Centers, University of Minnesota. The flow cytometry in this work was carried out at the University of Minnesota Flow Cytometry Resource. The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program. Funding Information: This work was supported by the National Institutes of Health (R01EB023880, R25HL128372), National Science Foundation Graduate Research Fellowship (CON‐75851, Project 00074041) and ERC‐2016‐CoG 725034 (ERC Consolidator Grant Des.solve). The confocal light microscopy in this work was carried out using a Zeiss Scope with the assistance of Guillermo Marques at the University Imaging Centers, University of Minnesota. The flow cytometry in this work was carried out at the University of Minnesota Flow Cytometry Resource. The Raman spectroscopy in this work was carried out in the Characterization Facility, University of Minnesota, which receives partial support from the NSF through the MRSEC program. Publisher Copyright: {\textcopyright} 2020 American Institute of Chemical Engineers",

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Natural deep eutectic systems for nature-inspired cryopreservation of cells

Abstract

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