NIFE Video

NIFE - Making-of




Head of the Institute: Prof. Prof. h.c. Dr.-Ing. Birgit Glasmacher, M.Sc. Institute for Multiphase Processes, Leibniz University Hannover phone: +49.511.762.3828, mail:

Speaker of the Research Group: Oleksandr Gryshkov, PhD phone: +49.511.532.1342 | +49.511.762.3822, mail:

Keywords:Biobanking, cell, tissue and organ cryopreservation, controlled rate freezing, freeze-drying, directional freezing, RAMAN-AFM, control of ice formation, cryomicroscopy, (epigenetic) analysis of cryopreservation outcome

Main Research Topics

The research group is focused on the development of safe and efficient cryopreservation protocols as well as novel freezing and thawing equipment for long-term storage and biobanking of cells and tissues

The main research interests of the group include the following:

  • Investigation of optimal freezing and thawing parameters as well as novel low toxic, defined and xeno-free freezing conditions resulting in high viability of a cryopreserved material (clinically relevant cells, tissues and tissue-engineered constructs) and their long-term epigenetic stability
  • Engineering and validation of unique and application-specific cryopreservation equipment with programmable freezing and thawing rates
  • Development of methods and equipment for induction and active control of ice nucleation at specific temperatures including low voltage electro-freezing, power down, electroporation techniques (in cooperation with University of Ljubljana, Slovenia)
  • Development of strategies for the visualization and analysis of ice formation and recrystallization to investigate cell and tissue specific cryopreservation parameters for effective Biobanking (in cooperation with the Hebrew University of Jerusalem, Israel)
  • Microscopy based visualization and analysis of cell and tissue freeze-drying using Linkam FCDS196 cryostage (in cooperation with KNURE, Ukraine)
  • Analysis of biochemical changes occurred during freezing, thawing and freeze-drying using RAMAN microscopy (confocal RAMAN microscope alpha 300RA from WiTec)
  • Cell protection using alginate encapsulation
  • Rheological characterization of novel cryoprotective solutions (CPA) and their impact on cell behavior during pre-treatment, freezing, storage and thawing.


  1. F. Bajerski, A. Bürger, B. Glasmacher, E.R.J. Keller, K. Müller, K. Mühldorfer, M. Nagel, H. Rüdel, T. Müller, J. Schenkel, J. Overmann. Factors determining microbial colonization of liquid nitrogen storage tanks used for archiving biological samples. Appl Microbiol Biotechnol. 2020;104(1): 131-144.

  2. V. Mutsenko, S. Knaack, L. Lauterboeck, D. Tarusin, B. Sydykov, R. Cabiscol, D. Ivnev, J. Belikan, A. Beck, D. Dipresa, A. Lode, T.El. Khassawna, M. Kampschulte, R. Scharf, A.Yu. Petrenko, S. Korossis, W.F. Wolkers, M. Gelinsky, B. Glasmacher, O. Gryshkov. Effect of ‘in air’ freezing on post-thaw recovery of Callithrix jacchus mesenchymal stromal cells and properties of 3D collagen-hydroxyapatite scaffolds. Cryobiology 2020, in press, doi: 10.1016/j.cryobiol.2020.01.015.

  3. V. Mutsenko, A. Barlič, T. Pezić, J. Dermol-Černe, B. Sydykov, W.F. Wolkers, I. Katkov, D. Miklavčič, B. Glasmacher, O. Gryshkov. Me2SO- and serum-free cryopreservation of mesenchymal stromal cells using electroporation of sugars. Cryobiology 2019;91: 104-114.

  4. O. Gryshkov, M. Müller, S. Leal-Marin, V. Mutsenko, S. Suresh, V.M. Kapralova, B. Glasmacher. Advances in the application of electrohydrodynamic fabrication for tissue engineering. J. Phys. Conf. Ser. 2019;1236 012024, doi 10.1088/1742-6596/1236/1/012024.

  5. V. Mutsenko, O. Gryshkov, O. Rogulska, A. Lode, A.Yu. Petrenko, M. Gelinsky, B. Glasmacher, H. Ehrlich. Chitinous Scaffolds from Marine Sponges for Tissue Engineering. In: A.H. Choi, B. Ben-Nissan (eds.) (2018). Marine-Derived Biomaterials for Tissue Engineering Applications, Springer Series in Biomaterials Science and Engineering (SSBSE) 2019: 285-307, doi 10.1007/978-981-13-8855-2_13.

  6. Patent DE 10 2018 100 844 (2019, pending), Process for freezing of biological cells and/or tissue in a fluid.

  7. M.V. Prykhodko, M.Y. Tymkovych, O.G. Avrunin, V.V. Mutsenko, O. Gryshkov, B. Glasmacher. Image processing for automated microscopic analysis of ice recrystallization process during isothermal annealing. Int J Bioelectromagnetism 2018;20(1): 72-75.

  8. M.Yu. Tymkovych, O.G. Avrunin, V.G. Paliy, M. Filzow, O. Gryshkov, B. Glasmacher et al. Automated method for structural segmentation of nasal airways based on cone beam assisted computed tomography. Proceedings of SPIE, Chapter: 10445, Editors: Romaniuk Ryszard S., Linczuk Maciej, pp.104453F, doi: 10.1117/12.2280922.

  9. V.M. Mutsenko. Cryopreservation of mesenchymal stromal cells within tissue engineering approaches (2019). PhD thesis, Hannover Medical School, Hannover.

  10. V.V. Mutsenko, V.V. Bazhenov, O. Rogulska, D.N. Tarusin, K. Schütz, S. Brüggemeier et al. 3D chitinous scaffolds derived from cultivated marine demosponge Aplysina aerophoba for tissue engineering approaches based on human mesenchymal stromal cells. Int J Biol Macromol. 2017;104(B): 1966-1974.

  11. V.V. Mutsenko, O. Gryshkov, L. Lauterboeck, O. Rogulska, D.N. Tarusin, V.V. Bazhenov et al. Novel chitin scaffolds derived from marine sponge Ianthella basta for tissue engineering approaches based on human mesenchymal stromal cells: Biocompatibility and cryopreservation. Int J Biol Macromol. 2017;104(B): 1955-1965.

  12. A. Chatterjee, D. Saha, H. Niemann, O. Gryshkov, B. Glasmacher, N. Hofmann. Effects of cryopreservation on the epigenetic profile of cells. Cryobiology 2017;74: 1-7.

  13. L. Lauterboeck, W.F. Wolkers, B. Glasmacher. Cryobiological parameters of multipotent stromal cells obtained from different sources. Cryobiology 2017;74: 93-102.

  14. D. Pogozhykh, Y. Pakhomova, O. Pervushina, N. Hofmann, B. Glasmacher, G. Zhegunov. Exploring the Possibility of Cryopreservation of Feline and Canine Erythrocytes by Rapid Freezing with Penetrating and NonPenetrating Cryoprotectants. PLoS ONE 2017;12(1): e0169689, DOI: 10.1371/journal.pone.0169689.

  15. L. Lauterboeck, D. Saha, A. Chatterjee, N. Hofmann, B. Glasmacher. Xeno-free cryopreservation of bone marrow derived multipotent stromal cells from Callithrix jacchus. Biopreserv Biobank. 2016;4(6): 530-538.

  16. A. Chatterjee, D. Saha, B. Glasmacher, N. Hofmann. Chilling without regrets: Deciphering the effects of cryopreservation on the epigenetic properties of frozen cells will benefit the applications of cryotechnology. EMBO Reports 2016;17: 292-295.

  17. A. Chatterjee. Effects of cryopreservation on histone posttranslational modifications of stem cells (2016). PhD Thesis, Hannover Medical School, Hannover.

  18. D. Saha. Effect of cryopreservation procedures on the viability, genetic and epigenetic stability of multipotent stromal cells (2016). PhD thesis, Hannover Medical School, Hannover.

  19. L. Lauterböck. Cryopreservation of stem cells using induced nucleation (2016). PhD thesis Hannover Medical School, Hannover.

  20. A. Repanas, L. Lauterboeck, D. Marvilas, B. Glasmacher. Polycaprolactone and polycaprolactone/ chitosan electrospun scaffolds for tissue engineering applications. Sch J App Med Sci. 2016:4(1C): 228-232.

  21. O. Gryshkov. High voltage encapsulation of multipotent stromal cells in alginate (2015). PhD thesis, Hannover Medical School, Hannover.

  22. O. Gryshkov, N. Hofmann, L. Lauterboeck, D. Pogozhykh, T. Mueller, B. Glasmacher. Multipotent Stromal Cells Derived from Common Marmoset Callithrix Jacchus within Alginate 3D Environment: Effect of Cryopreservation Procedures. Cryobiology 2015:71(1): 103-111.

  23. L. Lauterboeck, N. Hofmann, T. Mueller, B. Glasmacher. Active control of the nucleation temperature enhances freezing survival of multipotent mesenchymal stromal cells. Cryobiology 2015;17(3): 384-390.

  24. N. Hofmann, H. Sun, A. Chatterjee, D. Saha, B. Glasmacher. Thermal Pretreatment Improves Viability of Cryopreserved Human Endothelial Cells. Biopreserv Biobank. 2015;13: 348-355.

  25. O. Gryshkov, D. Pogozhykh, N. Hofmann, O. Pogozhykh, T. Mueller, B. Glasmacher. Encapsulating Non-Human Primate Multipotent Stromal Cells in Alginate via High Voltage for Cell-Based Therapies and Cryopreservation. PLoS One 2014;9(9): e107911.

  26. O. Gryshkov, D. Pogozhykh, N. Hofmann, N. Hofmann, T. Mueller, B. Glasmacher. Process engineering of high voltage alginate encapsulation of mesenchymal stem cells. Mater Sci Eng C Mater Biol Appl. 2014;36: 77-83.