Details
| Original language | English |
|---|---|
| Title of host publication | 8th European Medical and Biological Engineering Conference - Proceedings of the EMBEC 2020 |
| Editors | Tomaz Jarm, Aleksandra Cvetkoska, Samo Mahnič-Kalamiza, Damijan Miklavcic |
| Place of Publication | Cham |
| Publisher | Springer Science and Business Media Deutschland GmbH |
| Pages | 391-398 |
| Number of pages | 8 |
| ISBN (electronic) | 978-3-030-64610-3 |
| ISBN (print) | 9783030646097 |
| Publication status | Published - 30 Nov 2020 |
| Event | 8th European Medical and Biological Engineering Conference, EMBEC 2020 - Portorož, Slovenia Duration: 29 Nov 2020 → 3 Dec 2020 |
Publication series
| Name | IFMBE Proceedings |
|---|---|
| Volume | 80 |
| ISSN (Print) | 1680-0737 |
| ISSN (electronic) | 1433-9277 |
Abstract
As novel tissue engineered constructs (TECs) are developed, current tissue banking practices need better control over ice formation and growth to prevent cryodamage to cells and a scaffold. Directional solidification demonstrates benefits in adhered cells and native tissues cryopreservation through controlled heat transfer. Therefore, this study aims to investigate the feasibility of using this technique for cryopreservation of cell-seeded electrospun fiber mats as model TECs. Fiber mats were produced using blend electrospinning of polycaprolactone (PCL, 200 mg/ml) and poly-L-lactic acid (PLA, 100 mg/ml) dissolved in 2,2,2-Trifluoroethanol. The fiber size and morphology was characterized using scanning electron microscopy. Specific heat measurements were conducted using differential scanning calorimetry. The square-shaped fiber mats were seeded under static conditions with HeLa cells and cultivated for 24 h. Samples were directionally frozen in a sandwich format either in 10% DMSO or culture medium with the sample movement at 30 μm/s through the predetermined temperature gradients along a 2.6 mm slit. After directional solidification, samples were gradually frozen at 1 K/min down to −80 ℃. Crystal shape was visualized using cryomicroscopic system. Before freezing and 24 h after thawing, cell viability was assessed using live-dead assay. Within randomly orientated PCL-PLA fibers, HeLa cells exhibited typical shape and attachment with higher than 90% viability prior to freezing. While up to 80% of HeLa cells were alive on fiber mats after freezing using DMSO with or without directional solidification step. The demonstrated controlled freezing may assist optimizing the freezing of more sensitive cells. The results suggest that directional freezing becomes a viable option for cryopreservation in tissue engineering applications.
Keywords
- Cryopreservation, Directional freezing, Electrospun fiber mats
ASJC Scopus subject areas
- Chemical Engineering(all)
- Bioengineering
- Engineering(all)
- Biomedical Engineering
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8th European Medical and Biological Engineering Conference - Proceedings of the EMBEC 2020. ed. / Tomaz Jarm; Aleksandra Cvetkoska; Samo Mahnič-Kalamiza; Damijan Miklavcic. Cham: Springer Science and Business Media Deutschland GmbH, 2020. p. 391-398 (IFMBE Proceedings; Vol. 80).
Research output: Chapter in book/report/conference proceeding › Conference contribution › Research › peer review
}
TY - GEN
T1 - Directional Freezing of Cell-Seeded Electrospun Fiber Mats for Tissue Engineering Applications
AU - Mutsenko, Vitalii
AU - Chasnitsky, Michael
AU - Sirotinskaya, Vera
AU - Müller, Marc
AU - Glasmacher, Birgit
AU - Braslavsky, Ido
AU - Gryshkov, Oleksandr
N1 - Funding Information: Acknowledgements. This work was sponsored by Short-Term Research Grant from Minerva Foundation, the Israel Science Foundation (grant number 930/16) as well as Ways to Research II Program of the Leibniz University Hannover (60442522).
PY - 2020/11/30
Y1 - 2020/11/30
N2 - As novel tissue engineered constructs (TECs) are developed, current tissue banking practices need better control over ice formation and growth to prevent cryodamage to cells and a scaffold. Directional solidification demonstrates benefits in adhered cells and native tissues cryopreservation through controlled heat transfer. Therefore, this study aims to investigate the feasibility of using this technique for cryopreservation of cell-seeded electrospun fiber mats as model TECs. Fiber mats were produced using blend electrospinning of polycaprolactone (PCL, 200 mg/ml) and poly-L-lactic acid (PLA, 100 mg/ml) dissolved in 2,2,2-Trifluoroethanol. The fiber size and morphology was characterized using scanning electron microscopy. Specific heat measurements were conducted using differential scanning calorimetry. The square-shaped fiber mats were seeded under static conditions with HeLa cells and cultivated for 24 h. Samples were directionally frozen in a sandwich format either in 10% DMSO or culture medium with the sample movement at 30 μm/s through the predetermined temperature gradients along a 2.6 mm slit. After directional solidification, samples were gradually frozen at 1 K/min down to −80 ℃. Crystal shape was visualized using cryomicroscopic system. Before freezing and 24 h after thawing, cell viability was assessed using live-dead assay. Within randomly orientated PCL-PLA fibers, HeLa cells exhibited typical shape and attachment with higher than 90% viability prior to freezing. While up to 80% of HeLa cells were alive on fiber mats after freezing using DMSO with or without directional solidification step. The demonstrated controlled freezing may assist optimizing the freezing of more sensitive cells. The results suggest that directional freezing becomes a viable option for cryopreservation in tissue engineering applications.
AB - As novel tissue engineered constructs (TECs) are developed, current tissue banking practices need better control over ice formation and growth to prevent cryodamage to cells and a scaffold. Directional solidification demonstrates benefits in adhered cells and native tissues cryopreservation through controlled heat transfer. Therefore, this study aims to investigate the feasibility of using this technique for cryopreservation of cell-seeded electrospun fiber mats as model TECs. Fiber mats were produced using blend electrospinning of polycaprolactone (PCL, 200 mg/ml) and poly-L-lactic acid (PLA, 100 mg/ml) dissolved in 2,2,2-Trifluoroethanol. The fiber size and morphology was characterized using scanning electron microscopy. Specific heat measurements were conducted using differential scanning calorimetry. The square-shaped fiber mats were seeded under static conditions with HeLa cells and cultivated for 24 h. Samples were directionally frozen in a sandwich format either in 10% DMSO or culture medium with the sample movement at 30 μm/s through the predetermined temperature gradients along a 2.6 mm slit. After directional solidification, samples were gradually frozen at 1 K/min down to −80 ℃. Crystal shape was visualized using cryomicroscopic system. Before freezing and 24 h after thawing, cell viability was assessed using live-dead assay. Within randomly orientated PCL-PLA fibers, HeLa cells exhibited typical shape and attachment with higher than 90% viability prior to freezing. While up to 80% of HeLa cells were alive on fiber mats after freezing using DMSO with or without directional solidification step. The demonstrated controlled freezing may assist optimizing the freezing of more sensitive cells. The results suggest that directional freezing becomes a viable option for cryopreservation in tissue engineering applications.
KW - Cryopreservation
KW - Directional freezing
KW - Electrospun fiber mats
UR - http://www.scopus.com/inward/record.url?scp=85097615407&partnerID=8YFLogxK
U2 - 10.1007/978-3-030-64610-3_45
DO - 10.1007/978-3-030-64610-3_45
M3 - Conference contribution
AN - SCOPUS:85097615407
SN - 9783030646097
T3 - IFMBE Proceedings
SP - 391
EP - 398
BT - 8th European Medical and Biological Engineering Conference - Proceedings of the EMBEC 2020
A2 - Jarm, Tomaz
A2 - Cvetkoska, Aleksandra
A2 - Mahnič-Kalamiza, Samo
A2 - Miklavcic, Damijan
PB - Springer Science and Business Media Deutschland GmbH
CY - Cham
T2 - 8th European Medical and Biological Engineering Conference, EMBEC 2020
Y2 - 29 November 2020 through 3 December 2020
ER -