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

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Vitalii Mutsenko - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Sven Knaack - , Medizinische Fakultät Carl Gustav Carus Dresden (Autor:in)
  • Lothar Lauterboeck - , Louisiana State University Health Sciences Center (Autor:in)
  • Dmytro Tarusin - , NASU - Institute for Problems of Cryobiology and Cryomedicine (Autor:in)
  • Bulat Sydykov - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Ramon Cabiscol - , Technische Universität Braunschweig (Autor:in)
  • Dmitrii Ivnev - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Jan Belikan - , Justus-Liebig-Universität Gießen (Autor:in)
  • Annemarie Beck - , Medizinische Hochschule Hannover (MHH) (Autor:in)
  • Daniele Dipresa - , Medizinische Hochschule Hannover (MHH) (Autor:in)
  • Anja Lode - , Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung (Autor:in)
  • Thaqif El Khassawna - , Justus-Liebig-Universität Gießen (Autor:in)
  • Marian Kampschulte - , Justus-Liebig-Universität Gießen (Autor:in)
  • Roland Scharf - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Alexander Yu Petrenko - , NASU - Institute for Problems of Cryobiology and Cryomedicine (Autor:in)
  • Sotirios Korossis - , Medizinische Hochschule Hannover (MHH), Loughborough University (Autor:in)
  • Willem F. Wolkers - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Michael Gelinsky - , Zentrum für Translationale Knochen-, Gelenk- und Weichgewebeforschung (Autor:in)
  • Birgit Glasmacher - , Leibniz Universität Hannover (LUH) (Autor:in)
  • Oleksandr Gryshkov - , Leibniz Universität Hannover (LUH) (Autor:in)

Abstract

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.

Details

OriginalspracheEnglisch
Seiten (von - bis)215-230
Seitenumfang16
FachzeitschriftCryobiology : international journal of low temperature biology and medicine
Jahrgang92
PublikationsstatusVeröffentlicht - 1 Feb. 2020
Peer-Review-StatusJa

Externe IDs

PubMed 31972153
ORCID /0000-0001-9075-5121/work/160047961

Schlagworte

Schlagwörter

  • Mesenchymal stromal cells, Sucrose pretreatment, Tissue-engineered constructs, ‘In air’ freezing