Details
| Original language | English |
|---|---|
| Pages (from-to) | 215-230 |
| Number of pages | 16 |
| Journal | Cryobiology |
| Volume | 92 |
| Early online date | 20 Jan 2020 |
| Publication status | Published - 1 Feb 2020 |
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.
Keywords
- Mesenchymal stromal cells, Sucrose pretreatment, Tissue-engineered constructs, ‘In air’ freezing
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- General Biochemistry,Genetics and Molecular Biology
- Agricultural and Biological Sciences(all)
- General Agricultural and Biological Sciences
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In: Cryobiology, Vol. 92, 01.02.2020, p. 215-230.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Effect of ‘in air’ freezing on post-thaw recovery of Callithrix jacchus mesenchymal stromal cells and properties of 3D collagen-hydroxyapatite scaffolds
AU - Mutsenko, Vitalii
AU - Knaack, Sven
AU - Lauterboeck, Lothar
AU - Tarusin, Dmytro
AU - Sydykov, Bulat
AU - Cabiscol, Ramon
AU - Ivnev, Dmitrii
AU - Belikan, Jan
AU - Beck, Annemarie
AU - Dipresa, Daniele
AU - Lode, Anja
AU - El Khassawna, Thaqif
AU - Kampschulte, Marian
AU - Scharf, Roland
AU - Petrenko, Alexander Yu
AU - Korossis, Sotirios
AU - Wolkers, Willem F.
AU - Gelinsky, Michael
AU - Glasmacher, Birgit
AU - Gryshkov, Oleksandr
N1 - Funding Information: This work was supported by the German Research Foundation through the Cluster of Excellence REBIRTH (EXC 62/3 valid until Dec 2017, EXC 62/4 valid until Oct 2019), IP@Leibniz Program of the Leibniz University Hannover promoted by the German Academic Exchange Service (DAAD) (project code 57156199 ) as well as Ways to Research II Program of the Leibniz University Hannover ( 60442522 ).
PY - 2020/2/1
Y1 - 2020/2/1
N2 - 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.
AB - 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.
KW - Mesenchymal stromal cells
KW - Sucrose pretreatment
KW - Tissue-engineered constructs
KW - ‘In air’ freezing
UR - http://www.scopus.com/inward/record.url?scp=85078237703&partnerID=8YFLogxK
U2 - 10.1016/j.cryobiol.2020.01.015
DO - 10.1016/j.cryobiol.2020.01.015
M3 - Article
C2 - 31972153
AN - SCOPUS:85078237703
VL - 92
SP - 215
EP - 230
JO - Cryobiology
JF - Cryobiology
SN - 0011-2240
ER -