Directional Freezing of Cell-Seeded Electrospun Fiber Mats for Tissue Engineering Applications

Vitalii Mutsenko, Michael Chasnitsky, Vera Sirotinskaya, Marc Müller, Birgit Glasmacher, Ido Braslavsky, Oleksandr Gryshkov*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

2 Scopus citations


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.

Original languageAmerican English
Title of host publication8th European Medical and Biological Engineering Conference - Proceedings of the EMBEC 2020
EditorsTomaz Jarm, Aleksandra Cvetkoska, Samo Mahnič-Kalamiza, Damijan Miklavcic
PublisherSpringer Science and Business Media Deutschland GmbH
Number of pages8
ISBN (Print)9783030646097
StatePublished - 2021
Event8th European Medical and Biological Engineering Conference, EMBEC 2020 - Portorož, Slovenia
Duration: 29 Nov 20203 Dec 2020

Publication series

NameIFMBE Proceedings
ISSN (Print)1680-0737
ISSN (Electronic)1433-9277


Conference8th European Medical and Biological Engineering Conference, EMBEC 2020

Bibliographical note

Publisher Copyright:
© 2021, Springer Nature Switzerland AG.


  • Cryopreservation
  • Directional freezing
  • Electrospun fiber mats


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