TY - JOUR

T1 - Coaxial Alginate Hydrogels: From Self-Assembled 3D Cellular Constructs to Long-Term Storage

AU - Gryshkov, Oleksandr

AU - Mutsenko, Vitalii

AU - Tarusin, Dmytro

AU - Khayyat, Diaa

AU - Naujok, Ortwin

AU - Riabchenko, Ekaterina

AU - Nemirovska, Yuliia

AU - Danilov, Arseny

AU - Petrenko, Alexander Y.

AU - Glasmacher, Birgit

N1 - Funding Information: Acknowledgments: The authors would like to express their sincere grateful to Thomas Müller and Olena Pogozhykh (Institute for Transfusion Medicine, Hannover Medical School) for providing the stocks of cjaMSCs for the experiments as well as Annemarie Beck from the Department of Cardiothoracic, Transplantation and Vascular Surgery (Hannover Medical School, Hannover, Germany) for the assistance with the histological analysis. This work was technically supported by the students Yajarupika Kirupananthan, Jan Mairose and Björn Wünschmann. Authors are especially grateful to Katerina Zelena for her excellent technical support as well as Sven-Alexander Barker for additional language editing during submission and revision process.

PY - 2021/3/18

Y1 - 2021/3/18

N2 - Alginate as a versatile naturally occurring biomaterial has found widespread use in the biomedical field due to its unique features such as biocompatibility and biodegradability. The ability of its semipermeable hydrogels to provide a favourable microenvironment for clinically relevant cells made alginate encapsulation a leading technology for immunoisolation, 3D culture, cryopreservation as well as cell and drug delivery. The aim of this work is the evaluation of structural properties and swelling behaviour of the core-shell capsules for the encapsulation of multipotent stromal cells (MSCs), their 3D culture and cryopreservation using slow freezing. The cells were encapsulated in core-shell capsules using coaxial electrospraying, cultured for 35 days and cryopreserved. Cell viability, metabolic activity and cell–cell interactions were analysed. Cryopreservation of MSCs-laden core-shell capsules was performed according to parameters pre-selected on cell-free capsules. The results suggest that core-shell capsules produced from the low viscosity high-G alginate are superior to high-M ones in terms of stability during in vitro culture, as well as to solid beads in terms of promoting formation of viable self-assembled cellular structures and maintenance of MSCs functionality on a long-term basis. The application of 0.3 M sucrose demonstrated a beneficial effect on the integrity of capsules and viability of formed 3D cell assemblies, as compared to 10% dimethyl sulfoxide (DMSO) alone. The proposed workflow from the preparation of core-shell capsules with self-assembled cellular structures to the cryopreservation appears to be a promising strategy for their off-the-shelf availability.

AB - Alginate as a versatile naturally occurring biomaterial has found widespread use in the biomedical field due to its unique features such as biocompatibility and biodegradability. The ability of its semipermeable hydrogels to provide a favourable microenvironment for clinically relevant cells made alginate encapsulation a leading technology for immunoisolation, 3D culture, cryopreservation as well as cell and drug delivery. The aim of this work is the evaluation of structural properties and swelling behaviour of the core-shell capsules for the encapsulation of multipotent stromal cells (MSCs), their 3D culture and cryopreservation using slow freezing. The cells were encapsulated in core-shell capsules using coaxial electrospraying, cultured for 35 days and cryopreserved. Cell viability, metabolic activity and cell–cell interactions were analysed. Cryopreservation of MSCs-laden core-shell capsules was performed according to parameters pre-selected on cell-free capsules. The results suggest that core-shell capsules produced from the low viscosity high-G alginate are superior to high-M ones in terms of stability during in vitro culture, as well as to solid beads in terms of promoting formation of viable self-assembled cellular structures and maintenance of MSCs functionality on a long-term basis. The application of 0.3 M sucrose demonstrated a beneficial effect on the integrity of capsules and viability of formed 3D cell assemblies, as compared to 10% dimethyl sulfoxide (DMSO) alone. The proposed workflow from the preparation of core-shell capsules with self-assembled cellular structures to the cryopreservation appears to be a promising strategy for their off-the-shelf availability.

KW - Cell encapsulation

KW - Cellular structures

KW - Coaxial electrospraying

KW - Core-shell capsules

KW - Multipotent stromal cells

KW - RAMAN spec-troscopy

KW - Scaffolds

KW - Swelling

KW - Thermomechanical stress

KW - Tissue cryopreservation

UR - http://www.scopus.com/inward/record.url?scp=85102625023&partnerID=8YFLogxK

U2 - 10.3390/ijms22063096

DO - 10.3390/ijms22063096

M3 - Article

C2 - 33803546

AN - SCOPUS:85102625023

VL - 22

SP - 1

EP - 31

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1661-6596

IS - 6

M1 - 3096

ER -