Details
| Originalsprache | Englisch |
|---|---|
| Titel des Sammelwerks | 8th European Medical and Biological Engineering Conference |
| Untertitel | Proceedings of the EMBEC 2020 |
| Herausgeber/-innen | Tomaz Jarm, Aleksandra Cvetkoska, Samo Mahnič-Kalamiza, Damijan Miklavcic |
| Herausgeber (Verlag) | Springer Science and Business Media Deutschland GmbH |
| Seiten | 485-494 |
| Seitenumfang | 10 |
| ISBN (elektronisch) | 978-3-030-64610-3 |
| ISBN (Print) | 9783030646097 |
| Publikationsstatus | Veröffentlicht - 30 Nov. 2020 |
| Veranstaltung | 8th European Medical and Biological Engineering Conference, EMBEC 2020 - Portorož, Slowenien Dauer: 29 Nov. 2020 → 3 Dez. 2020 |
Publikationsreihe
| Name | IFMBE Proceedings |
|---|---|
| Band | 80 |
| ISSN (Print) | 1680-0737 |
| ISSN (elektronisch) | 1433-9277 |
Abstract
We have recently reported on the high practical utility of using electroporation of clinically relevant cells with sugars for their xeno-free cryopreservation in suspension. This paper extends our earlier approach to in situ electroporating attached cells for their robust cryopreservation on artificial scaffolds. Using CAD modelling, a two-electrode setup has been designed and in-house constructed allowing for simultaneous electroporation in multi-well cell culture plates. Blend electrospinning process has been optimized in order to manufacture porous fibrous mats made of polycaprolactone (PCL, 100 mg/ml) and polylactide (PLA, 50 mg/ml) with an average thickness of 100 µm. Chinese hamster ovary (CHO) cells were grown and directly electroporated in nanofibrous blend electrospun fiber mats. An electric pulse was applied in the presence of propidium iodide and CellTracker Green to determine viable permeabilized cell counts using fluorescence microscopy. Cell recovery was evaluated using metabolic MTS assay 24 h post-electroporation. Electric field intensity and distribution within a 3D reconstructed fiber mat was simulated and visualized using COMSOL software. The results demonstrate that with developed setup it is feasible to electroporate around 80% of attached cells with 80% viability after electroporation when electric field strength was ≥1.7 kV/cm. COMSOL simulations showed local increases of electric field at intersection points of numerous fibers which may in part contribute to the observed drop in cell viability post-electroporation. Future studies anticipate implementation of the developed approach in effective biopreservation of stem cells on electrospun fiber mats as a model of tissue-engineered constructs.
ASJC Scopus Sachgebiete
- Chemische Verfahrenstechnik (insg.)
- Bioengineering
- Ingenieurwesen (insg.)
- Biomedizintechnik
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8th European Medical and Biological Engineering Conference: Proceedings of the EMBEC 2020. Hrsg. / Tomaz Jarm; Aleksandra Cvetkoska; Samo Mahnič-Kalamiza; Damijan Miklavcic. Springer Science and Business Media Deutschland GmbH, 2020. S. 485-494 (IFMBE Proceedings; Band 80).
Publikation: Beitrag in Buch/Bericht/Sammelwerk/Konferenzband › Aufsatz in Konferenzband › Forschung › Peer-Review
}
TY - GEN
T1 - Electroporation of Cell-Seeded Electrospun Fiber Mats for Cryopreservation
AU - Gryshkov, Oleksandr
AU - Mutsenko, Vitalii
AU - Dermol-Černe, Janja
AU - Miklavčič, Damijan
AU - Glasmacher, Birgit
N1 - Funding Information: Acknowledgements. This work was in part performed in the framework of student exchange by Tobias Pfister whose stay at the University of Ljubljana was sponsored by the Dr. Jürgen and Irmgard Ulderup Foundation. The authors acknowledge technical assistance of the student.
PY - 2020/11/30
Y1 - 2020/11/30
N2 - We have recently reported on the high practical utility of using electroporation of clinically relevant cells with sugars for their xeno-free cryopreservation in suspension. This paper extends our earlier approach to in situ electroporating attached cells for their robust cryopreservation on artificial scaffolds. Using CAD modelling, a two-electrode setup has been designed and in-house constructed allowing for simultaneous electroporation in multi-well cell culture plates. Blend electrospinning process has been optimized in order to manufacture porous fibrous mats made of polycaprolactone (PCL, 100 mg/ml) and polylactide (PLA, 50 mg/ml) with an average thickness of 100 µm. Chinese hamster ovary (CHO) cells were grown and directly electroporated in nanofibrous blend electrospun fiber mats. An electric pulse was applied in the presence of propidium iodide and CellTracker Green to determine viable permeabilized cell counts using fluorescence microscopy. Cell recovery was evaluated using metabolic MTS assay 24 h post-electroporation. Electric field intensity and distribution within a 3D reconstructed fiber mat was simulated and visualized using COMSOL software. The results demonstrate that with developed setup it is feasible to electroporate around 80% of attached cells with 80% viability after electroporation when electric field strength was ≥1.7 kV/cm. COMSOL simulations showed local increases of electric field at intersection points of numerous fibers which may in part contribute to the observed drop in cell viability post-electroporation. Future studies anticipate implementation of the developed approach in effective biopreservation of stem cells on electrospun fiber mats as a model of tissue-engineered constructs.
AB - We have recently reported on the high practical utility of using electroporation of clinically relevant cells with sugars for their xeno-free cryopreservation in suspension. This paper extends our earlier approach to in situ electroporating attached cells for their robust cryopreservation on artificial scaffolds. Using CAD modelling, a two-electrode setup has been designed and in-house constructed allowing for simultaneous electroporation in multi-well cell culture plates. Blend electrospinning process has been optimized in order to manufacture porous fibrous mats made of polycaprolactone (PCL, 100 mg/ml) and polylactide (PLA, 50 mg/ml) with an average thickness of 100 µm. Chinese hamster ovary (CHO) cells were grown and directly electroporated in nanofibrous blend electrospun fiber mats. An electric pulse was applied in the presence of propidium iodide and CellTracker Green to determine viable permeabilized cell counts using fluorescence microscopy. Cell recovery was evaluated using metabolic MTS assay 24 h post-electroporation. Electric field intensity and distribution within a 3D reconstructed fiber mat was simulated and visualized using COMSOL software. The results demonstrate that with developed setup it is feasible to electroporate around 80% of attached cells with 80% viability after electroporation when electric field strength was ≥1.7 kV/cm. COMSOL simulations showed local increases of electric field at intersection points of numerous fibers which may in part contribute to the observed drop in cell viability post-electroporation. Future studies anticipate implementation of the developed approach in effective biopreservation of stem cells on electrospun fiber mats as a model of tissue-engineered constructs.
KW - Cryopreservation
KW - Electropermeabilization
KW - Electroporation
KW - Electrospun fiber mats
KW - Tissue engineering
UR - http://www.scopus.com/inward/record.url?scp=85097608424&partnerID=8YFLogxK
U2 - 10.1007/978-3-030-64610-3_55
DO - 10.1007/978-3-030-64610-3_55
M3 - Conference contribution
AN - SCOPUS:85097608424
SN - 9783030646097
T3 - IFMBE Proceedings
SP - 485
EP - 494
BT - 8th European Medical and Biological Engineering Conference
A2 - Jarm, Tomaz
A2 - Cvetkoska, Aleksandra
A2 - Mahnič-Kalamiza, Samo
A2 - Miklavcic, Damijan
PB - Springer Science and Business Media Deutschland GmbH
T2 - 8th European Medical and Biological Engineering Conference, EMBEC 2020
Y2 - 29 November 2020 through 3 December 2020
ER -