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Cryopreservation of mesenchymal stem cells using medical grade ice nucleation inducer

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posted on 24.11.2020, 11:20 by Nicholas M Wragg, Dimitris Tampakis, Alexandra StolzingAlexandra Stolzing
Mesenchymal stem cells (MSCs) can differentiate into multiple different tissue lineages and have favourable immunogenic potential making them an attractive prospect for regenerative medicine. As an essential part of the manufacturing process, preservation of these cells whilst maintaining potential is of critical importance. An uncontrolled area of storage remains the rate of change of temperature during freezing and thawing. Controlled-rate freezers attempted to rectify this; however, the change of phase from liquid to solid introduces two extreme phenomena; a rapid rise and a rapid fall in temperature in addition to the intended cooling rate (normally -1 °C/min) as a part of the supercooling event in cryopreservation. Nucleation events are well known to initiate the freezing transition although their active use in the form of ice nucleation devices (IND) are in their infancy in cryopreservation. This study sought to better understand the effects of ice nucleation and its active instigation with the use of an IND in both a standard cryotube with MSCs in suspension and a high-throughput adhered MSC 96-well plate set-up. A potential threshold nucleation temperature for best recovery of dental pulp MSCs may occur around -10 °C and for larger volume cell storage, IND and fast thaw creates the most stable process. For adhered cells, an IND with a slow thaw enables greatest metabolic activity post-thaw. This demonstrates a necessity for a medical grade IND to be used in future regenerative medicine manufacturing with the parameters discussed in this study to create stable products for clinical cellular therapies.

Funding

European Union’s Horizon 2020 research and innovation programme under grant agreement No. 667421

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

International Journal of Molecular Sciences

Volume

21

Issue

22

Publisher

MDPI

Version

VoR (Version of Record)

Rights holder

© The Authors

Publisher statement

This is an Open Access Article. It is published by MDPI under the Creative Commons Attribution 4.0 International Licence (CC BY 4.0). Full details of this licence are available at: https://creativecommons.org/licenses/by/4.0/

Acceptance date

29/10/2020

Publication date

2020-11-13

Copyright date

2020

ISSN

1661-6596

eISSN

1422-0067

Language

en

Depositor

Prof Alexandra Stolzing. Deposit date: 23 November 2020

Article number

8579

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