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Effect of ‘in air’ freezing on post-thaw recovery of Callithrix jacchus mesenchymal stromal cells and properties of 3D collagen-hydroxyapatite scaffolds

journal contribution
posted on 25.11.2020, 14:01 by Vitalii Mutsenko, Sven Knaack, Lothar Lauterboeck, Dmytro Tarusin, Bulat Sydykov, Ramon Cabiscol, Dmitrii Ivnev, Jan Belikan, Annemarie Beck, Daniele Dipresa, Anja Lode, Thaqif El Khassawna, Marian Kampschulte, Roland Scharf, Alexander Yu. Petrenko, Sotiris KorossisSotiris Korossis, Willem F. Wolkers, Michael Gelinsky, Birgit Glasmacher, Oleksandr Gryshkov
Through enabling an efficient supply of cells and tissues in the health sector on demand, cryopreservation is increasingly becoming one of the mainstream technologies in rapid translation and commercialization of regenerative medicine research. Cryopreservation of tissue-engineered constructs (TECs) is an emerging trend that requires the development of practically competitive biobanking technologies. In our previous studies, we demonstrated that conventional slow-freezing using dimethyl sulfoxide (Me2SO) does not provide sufficient protection of mesenchymal stromal cells (MSCs) frozen in 3D collagen-hydroxyapatite scaffolds. After simple modifications to a cryopreservation protocol, we report on significantly improved cryopreservation of TECs. Porous 3D scaffolds were fabricated using freeze-drying of a mineralized collagen suspension and following chemical crosslinking. Amnion-derived MSCs from common marmoset monkey Callithrix jacchus were seeded onto scaffolds in static conditions. Cell-seeded scaffolds were subjected to 24 h pre-treatment with 100 mM sucrose and slow freezing in 10% Me2SO/20% FBS alone or supplemented with 300 mM sucrose. Scaffolds were frozen ‘in air’ and thawed using a two-step procedure. Diverse analytical methods were used for the interpretation of cryopreservation outcome for both cell-seeded and cell-free scaffolds. In both groups, cells exhibited their typical shape and well-preserved cell-cell and cell-matrix contacts after thawing. Moreover, viability test 24 h post-thaw demonstrated that application of sucrose in the cryoprotective solution preserves a significantly greater portion of sucrose-pretreated cells (more than 80%) in comparison to Me2SO alone (60%). No differences in overall protein structure and porosity of frozen scaffolds were revealed whereas their compressive stress was lower than in the control group. In conclusion, this approach holds promise for the cryopreservation of ‘ready-to-use’ TECs.

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

Cryobiology

Volume

92

Pages

215 - 230

Publisher

Elsevier

Version

VoR (Version of Record)

Rights holder

© Elsevier

Publisher statement

This paper was accepted for publication in the journal Cryobiology and the definitive published version is available at https://doi.org/10.1016/j.cryobiol.2020.01.015.

Acceptance date

17/01/2020

Publication date

2020-01-20

Copyright date

2020

ISSN

0011-2240

eISSN

1090-2392

Language

en

Depositor

Prof Sotiris Korossis. Deposit date: 24 November 2020