Awards

  • 2016: Peter L. Steponkus Crystal Award for best oral presentation and student travel award for Miao Zhang, annual meeting of the Society for Cryobiology, Ottawa, Canada
  • 2015: John K. Critser Award for best abstract and student travel award for Miao Zhang, annual meeting of the Society for Cryobiology, Ostrava, Czech Republic

Biostabilization of tissues and macromolecular assemblies

Objectives

Biological materials used in regenerative medicine ultimately require preservation for successful translation in the clinical practice or for disease diagnostics. The main objective is to develop custom-designed preservation methods for bodily fluids, therapeutic cells, cell-seeded tissue-engineered constructs, and biological scaffolds for off-the-shelf availability. Examples include freeze-dried tissue-engineered heart valves, cryopreserved endothelialized oxygenators, direct transfusable cryopreserved therapeutic stem cells, dried shelf-stable bodily fluids (blood, saliva), and liposomal drug delivery systems.

Research Focus

The central research theme of the group is to study structure and stability of macromolecules in cells and tissues during extreme temperature and dehydration conditions. This encompasses development of mass transport models for protective agents and water into cells and tissues in order to develop preservation technology. Cells of interest include red blood cells, platelets, human pulmonary microvascular endothelial cells (HPMEC-ST1.6R), umbilical cord stem cells, mesenchymal stem cells, epithelial cells (293T), murine fibroblasts (NIH 3T3). Tissues of interest include decellularized heart valves and pericardium. Membrane transport parameters of cells are studied in microfluidic devices to predict optimal protocols to load cells with protective agents. Uptake methods to load cells with membrane impermeable trehalose are investigated for non-toxic cryopreservation and dry storage. Another main research theme focuses on diffusion studies to load decellularized heart valves with sugars so that they can be freeze-dried, which is moving towards translation. In collaboration with the Helmholtz Centre for Infection Research, a project aiming to implement a ‘Dry Biobank’ for bodily fluids (e.g. blood plasma) has been initiated to replace currently used cryopreservation approaches. Furthermore, dry preservation technology is being developed for saliva samples to support various large cohort studies on telomeres within the framework of rebirth-active.

Collaborations

  • Dr. Andres Hilfiker, Leibniz Research Laboratories for Biotechnology and Artificial Organs (LEBAO), Hannover Medical School, Hannover, Germany
  • Dr. Sotirios Korossis, Department of Cardiothoracic, Transplantation and Vascular Surgery, Hannover Medical School, Hannover, Germany
  • Prof. Dr. Uwe Tegtbur, Institute for Sport medicine, Hannover Medical School, Hannover, Germany
  • Dr. Eveline Sowa-Soehle, Biocompatibility Laboratory, Hannover Medical School, Hannover, Germany
  • Dr. Constanca Figueiredo, Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany
  • Dr. Tobias Kerrinnes, Helmholtz Centre for Infection Research, Braunschweig, Germany
  • Prof. Harald Sieme, Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany
  • Dr. Harriette Oldenhof, Unit for Reproductive Medicine - Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany
  • Dr. Michael Harder, corlife, Hannover, Germany Prof. Daniele Fioretto, Department of Physics, University of Perugia, Perugia, Italy
  • Dr. Laura Iop, University of Padua Medical School Dept. of Cardiac, Thoracic and Vascular Sciences, Padua, Italy
  • Prof. Dr. Adam Higgins, Oregon State University, Corvalis, OR, USA
  • Prof. Dr. John Bischof, Department of Mechanical Engineering, University of Minnesota, Minneapolis, MN, USA
  • Prof Fang Yang, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China
  • Prof. Dr. Jason Acker, Laboratory Medicine and Pathology, Edmonton, University of Alberta, Edmonton, Canada

Further Research Projects

  • Biomimetic liposomes
  • Tissue sterilization
  • Dry preservation of sperm

Teaching

  • Lecture and tutorial ‘Cryotechnology and cryobiology’
  • Lecture ‘Biomedizintechnik: freeze-drying’
  • Masterlabor ‘Biomedical process technology: liposome encapsulation and stabilization’
  • Tutorium ‘Bildgebung’
  • Lecture PhD program ‘regenerative sciences: tissue processing and stabilization’

Publications

2013 - ongoing

2018

Vásquez-Rivera A, Sommer KK, Oldenhof H, Higgins AZ, Brockbank KGM, Hilfiker A, Wolkers WF. Simultaneous monitoring of different vitrification solution components permeating into tissues. Analyst 2018;143:420-428.

Goecke T, Theodoridis K, Tudorache I, Ciubotaru A, Cebotari S, Ramm R, Höffler K, Sarikouch S, Vásquez Rivera A, Haverich A, Wolkers WF, Hilfiker A. In vivo performance of freeze-dried decellularized pulmonary heart valve allo- and xenografts orthotopically implanted into juvenile sheep. Acta Biomaterialia. 2018;68:41-52.

Sydykov B, Oldenhof H, de Oliveira Barros L, Sieme H, Wolkers WF. Membrane permeabilization of phosphatidylcholine liposomes induced by cryopreservation and vitrification solutions. Biochimica et Biophysica Acta (BBA) - Biomembranes. 2018;1860(2):467-74.

2017

Zhang M, Oldenhof H, Sydykov B, Bigalk J, Sieme H, Wolkers WF. Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity. Scientific Reports. 2017;7(1):6198.

Oldenhof H, Zhang M, Narten K, Bigalk J, Sydykov B, Wolkers WF, Sieme H. Freezing-induced uptake of disaccharides for preservation of chromatin in freeze-dried stallion sperm during accelerated aging†. Biology of Reproduction. 2017;97(6):892-901.

Sydykov B, Oldenhof H, Sieme H, Wolkers WF. Hydrogen Bonding Interactions and Enthalpy Relaxation in Sugar/Protein Glasses. Journal of Pharmaceutical Sciences. 2017;106(3):761-9.

Oldenhof H, Bigalk J, Hettel C, de Oliveira Barros L, Sydykov B, Bajcsy ÁC, Sieme H, Wolkers WF. Stallion Sperm Cryopreservation Using Various Permeating Agents: Interplay Between Concentration and Cooling Rate. Biopreservation and Biobanking. 2017;15(5):422-31.

Zhang M, Oldenhof H, Sieme H, Wolkers WF. Combining endocytic and freezing-induced trehalose uptake for cryopreservation of mammalian cells. Biotechnology Progress. 2017;33(1):229-35.

Ertmer F, Oldenhof H, Schütze S, Rohn K, Wolkers WF, Sieme H. Induced sub-lethal oxidative damage affects osmotic tolerance and cryosurvival of spermatozoa. Reproduction, Fertility and Development. 2017;29(9):1739-50.

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

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

2016

Zhang M, Oldenhof H, Sieme H, Wolkers WF. Combining endocytic and freezing-induced trehalose uptake for cryopreservation of mammalian cells. Biotechnol Prog. 2016.

Zhang M, Oldenhof H, Sieme H, Wolkers WF. Freezing-induced uptake of trehalose into mammalian cells facilitates cryopreservation. Bba-Biomembranes. 2016;1858(6):1400-9.

Sydykov B, Oldenhof H, Sieme H, Wolkers WF. Hydrogen Bonding Interactions and Enthalpy Relaxation in Sugar/Protein Glasses. J Pharm Sci. 2016.

Oldenhof H, Schutze S, Wolkers WF, Sieme H. Fourier transform infrared spectroscopic analysis of sperm chromatin structure and DNA stability. Andrology. 2016;4(3):430-41.

2015

Repanas A, Wolkers WF, Gryshkov O, Müller M, Glasmacher B. Pcl/Peg Electrospun fibers as drug carriers for the controlled delivery of dipyridamole. Journal of In Silico & In Vitro Pharmacology 1, 1:2, 10p. 2015.

Wang S. Characterization of diffusion processes for preservation of biological heart valve scaffolds. , PhD dissertation. Defense 15 Jan 2015.

Oldenhof H, Heutelbeck A, Blasse AK, Bollwein H, Martinsson G, Wolkers WF, Sieme H. Tolerance of Spermatozoa to Hypotonic Stress: Role of Membrane Fluidity and Correlation with Cryosurvival. Reprod Fertil Dev. 2015;27(2):285-93.

Sassi P, Caponi S, Ricci M, Morresi A, Oldenhof H, Wolkers WF, Fioretto D. Infrared Versus Light Scattering Techniques to Monitor the Gel to Liquid Crystal Phase Transition in Lipid Membranes. Journal of Raman Spectroscopy. 2015;46(7):644-51.

Sieme H, Oldenhof H, Wolkers WF. Sperm Membrane Behaviour During Cooling and Cryopreservation. Reprod Domest Anim. 2015;50 Suppl 3:20-6.

Wang S, Oldenhof H, Goecke T, Ramm R, Harder M, Haverich A, Hilfiker A, Wolkers WF. Sucrose Diffusion in Decellularized Heart Valves for Freeze-Drying. Tissue Eng Part C Methods. 2015;21(9):922-31.

Wolkers WF, Hilfiker A. Freeze-Drying of Decellularized Heart Valve Tissues. Methods Mol Biol. 2015;1257:499-506.

Wolkers WF, Oldenhof H. Use of in Situ Fourier Transform Infrared Spectroscopy to Study Freezing and Drying of Cells. Methods Mol Biol. 2015;1257:147-61.

Wolkers WF, Oldenhof H. Biomolecular fingerprinting in the dried state. Biomedical Spectroscopy and Imaging. 2015; 4: 331-340.

Repanas A, Wolkers WF, Gryshkov O, Kalozoumis P, Mueller M, Zernetsch H, Korossis S, Glasmacher B. Coaxial electrospinning as a process to engineer biodegradable polymeric scaffolds as a drug delivery systems for anti-inflammatory and anti-thrombotic pharmaceutical agents. Clinical & Experimental Pharmacology. 2015; 5:5, 8p.

2014

Qin Z, Balasubramanian SK, Wolkers WF, Pearce JA, Bischof JC. Correlated Parameter Fit of Arrhenius Model for Thermal Denaturation of Proteins and Cells. Ann Biomed Eng. 2014;42(12):2392-404.

Wang S, Oldenhof H, Dai X, Haverich A, Hilfiker A, Harder M, Wolkers WF. Protein Stability in Stored Decellularized Heart Valve Scaffolds and Diffusion Kinetics of Protective Molecules. Biochim Biophys Acta. 2014;1844(2):430-8.

2013

Oldenhof H, Gojowsky M, Wang S, Henke S, Yu C, Rohn K, Wolkers WF, Sieme H. Osmotic Stress and Membrane Phase Changes During Freezing of Stallion Sperm: Mode of Action of Cryoprotective Agents. Biol Reprod. 2013;88(3):68.

Schmid S, Henning H, Oldenhof H, Wolkers WF, Petrunkina AM, Waberski D. The Specific Response to Capacitating Stimuli Is a Sensitive Indicator of Chilling Injury in Hypothermically Stored Boar Spermatozoa. Andrology. 2013;1(3):376-86.

Oldenhof H, Akhoondi M, Sieme H, Wolkers WF. Use of Fourier transform infrared spectroscopy to determine optimal cooling rates for cryopreservation of cells. Biomedical Spectroscopy and Imaging. 2013;2(2):83-90.

Akhoondi, M. PhD dissertation: Modeling of water transport processes during freezing of mammalian cells. 15 Jan 2013.

2006 - 2012

2012

Akhoondi M, Oldenhof H, Sieme H, Wolkers WF. Freezing-Induced Cellular and Membrane Dehydration in the Presence of Cryoprotective Agents. Mol Membr Biol. 2012;29(6):197-206.

Blasse AK, Oldenhof H, Ekhlasi-Hundrieser M, Wolkers WF, Sieme H, Bollwein H. Osmotic Tolerance and Intracellular Ion Concentrations of Bovine Sperm Are Affected by Cryopreservation. Theriogenology. 2012;78(6):1312-20.

Glafke C, Akhoondi M, Oldenhof H, Sieme H, Wolkers WF. Cryopreservation of Platelets Using Trehalose: The Role of Membrane Phase Behavior During Freezing. Biotechnol Prog. 2012;28(5):1347-54.

Oldenhof H, Friedel K, Akhoondi M, Gojowsky M, Wolkers WF, Sieme H. Membrane Phase Behavior During Cooling of Stallion Sperm and Its Correlation with Freezability. Mol Membr Biol. 2012;29(3-4):95-106.

Sanchez V, Redmann K, Wistuba J, Wubbeling F, Burger M, Oldenhof H, Wolkers WF, Kliesch S, Schlatt S, Mallidis C. Oxidative DNA Damage in Human Sperm Can Be Detected by Raman Microspectroscopy. Fertil Steril. 2012;98(5):1124-9 e1-3.

Stoll C, Holovati JL, Acker JP, Wolkers WF. Synergistic Effects of Liposomes, Trehalose, and Hydroxyethyl Starch for Cryopreservation of Human Erythrocytes. Biotechnol Prog. 2012;28(2):364-71.

Wang S, Goecke T, Meixner C, Haverich A, Hilfiker A, Wolkers WF. Freeze-Dried Heart Valve Scaffolds. Tissue Eng Part C Methods. 2012;18(7):517-25.

Wang S, Oldenhof H, Hilfiker A, Harder M, Wolkers WF. Solvent accesibility of proteins in heart valve scaffolds. Biomedical Spectroscopy and Imaging 1 2012:79-87.

2011

Akhoondi M, Oldenhof H, Stoll C, Sieme H, Wolkers WF. Membrane Hydraulic Permeability Changes During Cooling of Mammalian Cells. Biochim Biophys Acta. 2011;1808(3):642-8.

Diekmann U, Spindler R, Wolkers WF, Glasmacher B, Muller T. Cryopreservation and Quality Control of Mouse Embryonic Feeder Cells. Cryobiology. 2011;63(2):104-10.

Oldenhof H, Blasse AK, Wolkers WF, Bollwein H, Sieme H. Osmotic Properties of Stallion Sperm Subpopulations Determined by Simultaneous Assessment of Cell Volume and Viability. Theriogenology. 2011;76(2):386-91.

Ropke T, Oldenhof H, Leiding C, Sieme H, Bollwein H, Wolkers WF. Liposomes for Cryopreservation of Bovine Sperm. Theriogenology. 2011;76(8):1465-72.

Spindler R, Wolkers WF, Glasmacher B. Dimethyl Sulfoxide and Ethylene Glycol Promote Membrane Phase Change During Cryopreservation. Cryo Letters. 2011;32(2):148-57.

Stadnick H, Stoll C, Wolkers WF, Acker JP, Holovati JL. The Effect of Liposome Treatment on the Quality of Hypothermically Stored Red Blood Cells. Biopreserv Biobank. 2011;9(4):335-42.

Stoll C, Holovati JL, Acker JP, Wolkers WF. Liposomes Composed of Unsaturated Lipids for Membrane Modification of Human Erythrocytes. Mol Membr Biol. 2011;28(7-8):454-61.

Stoll C, Stadnick H, Kollas O, Holovati JL, Glasmacher B, Acker JP, Wolkers WF. Liposomes Alter Thermal Phase Behavior and Composition of Red Blood Cell Membranes. Biochim Biophys Acta. 2011;1808(1):474-81.

Stoll C, Wolkers WF. Membrane Stability During Biopreservation of Blood Cells. Transfus Med Hemother. 2011;38(2):89-97.

2010

Venkatasubramanian RT, Wolkers WF, Shenoi MM, Barocas VH, Lafontaine D, Soule CL, Iaizzo PA, Bischof JC. Freeze-Thaw Induced Biomechanical Changes in Arteries: Role of Collagen Matrix and Smooth Muscle Cells. Ann Biomed Eng. 2010;38(3):694-706.

2009

Balasubramanian SK, Wolkers WF, Bischof JC. Membrane Hydration Correlates to Cellular Biophysics During Freezing in Mammalian Cells. Biochim Biophys Acta. 2009;1788(5):945-53.

Hagiwara M, Choi JH, Devireddy RV, Roberts KP, Wolkers WF, Makhlouf A, Bischof JC. Cellular Biophysics During Freezing of Rat and Mouse Sperm Predicts Post-Thaw Motility. Biol Reprod. 2009;81(4):700-6.

Shah NB, Wolkers WF, Morrissey M, Sun WQ, Bischof JC. Fourier Transform Infrared Spectroscopy Investigation of Native Tissue Matrix Modifications Using a Gamma Irradiation Process. Tissue Eng Part C Methods. 2009;15(1):33-40.

2007

Wolkers WF, Balasubramanian SK, Ongstad EL, Zec HC, Bischof JC. Effects of Freezing on Membranes and Proteins in Lncap Prostate Tumor Cells. Biochim Biophys Acta. 2007;1768(3):728-36.

Books edited:

Wolkers WF, Oldenhof H editors. Cryopreservation and Freeze-Drying Protocols. Book series: Methods in Molecular Biology, Vol 1275, New York, 528 p, ISBN 978-1-4939-2192-8.