1. Electroporation based Technologies and Treatments
Damijan Miklavčič
University of Ljubljana
Faculty of Electrical Engineering
Ljubljana, SLOVENIA

2. from the foreword to the
“Cells can be funny. Try to grow them with a slightly wrong recipe, and they turn over and die. But hit them with an electric field strong enough to knock over a horse, and they do enough things to justify international meetings, to fill a sizeable book, and to lead one to speak of an entirely new technology for cell manipulation.”
Adrian Parsegian
from the foreword to the
Electroporation and Electrofusion in Cell Biology
Edited by E. Neumann, A. S. Sowers and C. A. Jordan

4. Electroporation based Technologies and Treatments
Molecular cell biology research
Protein insertion into cell membrane
Cell fusion
Gene expression silencing by siRNA
Electrochemotherapy
Genetherapy based on electro genetransfer
Transdermal drug delivery
Tissue ablation
Biotechnology
Water and liquid food sterilisation

5. Electrochemotherapy = = chemotherapy + electroporation
Electric pulse generator
Chemotherapeutic surrounds the cells
Increased membrane permeability allows access to the cytosol
Pores reseal
Electrodes
time
Tumour
Electric pulse application
Injecting chemotherapeutic

6. Skin metastasis of squamous cell carcinoma of the supraglottis

7. Skin metastasis of malignant melanoma
Before treatment
One year after ECT

8. EU 5th FP: Quality of Life
Cliniporator QLK ( )
Electro-medical device for ECT
ESOPE QLK ( )
European Standard Operating Procedures for ECT

9. Objective Response Rate: 85% No Response: 15%
171 NODULES

10. GeneTherapy of diseases with altered angiogenesis:
Psoriasis and Malignant Melanoma

11. Electroporation of cells:
Experiments and modelling for better understanding

12. Cell (electro)fusion Monoclonal antibody production
Cancer immunotherapy
Receptor insertion into membrane
Tissue regeneration
Research (dissease models)

13. Dielectrophoresis V homogenem električnem polju ni dielektroforeze
Pozitivna dielektroforeza Negativna dielektroforeza
V homogenem električnem polju ni dielektroforeze
Dielectrophoresis

14. Biochip and results of experiments
CHO cellls
Medium conducivity 15 mS/cm
negativve dielectrofporesis
frequency 500 Hz
amplitude 4 V

15. Molecular cell biology research Electrochemotherapy Electrogenetherapy
Electroporation based Technologies and Treatments
Molecular cell biology research
Electrochemotherapy
Electrogenetherapy
Gene expression silencing by siRNA
Transdermal drug delivery
Protein insertion into cell membrane
Cell fusion
Biotechnology
Water and liquid food sterilisation
Tissue ablation

16. LBK expertise and resources:
Cell biology and chemistry lab
Lipid bilayer setup for preparation of planar lipid membranes.
chamber for lipid bilayers
microscope with cold-light illumination for lipid bilayer observation
(SZF 30, Olympus, Germany)

17. LBK expertise and resources:
Cell biology lab
Prototype of an electroporator developed in LBK connected to digital oscilloscope LeCroy 9310C.
Inverted phase contrast microscope Olympus CK40 for routine work with cell cultures & lipid vesicles.

18. LBK expertise and resources:
Cell biology lab
System for dynamic imaging of cells and lipid vesicles.
Zeiss Axiovert 200 epiflurescence
inverted microscope (Zeiss, Germany)
high resolution cooled CCD camera
(VisiCam1280)
sensitive high-speed cooled CCD
camera (BOOST 2000, Diagnostic
Instruments, GB)

19. List of other equipment
Other equipment available:
Spectrofluorometer (Jasco, Japan)
Microplate reader (Tecan, Switzerland)
Field jump system for fast monitoring optical changes in lipid vesicles
Computer facilities
Software: Mathematica, Matlab with different toolboxes;
Numerical modeling software: MSC/EMAS, Maxwell, COMSOL Multiphysics
NIRO2-X2: near infrared spectrophotometer for measurement of oxygenation and blood perfusion changes in tissue (Keele University, U.K.)
OxyLite 2000: 2-channel instrument for luminescence-based fiber-optic oximetry in tissue; OxyFlo 2000: 2-channel instrument for laser Doppler flowmetry in tissue; OxyData 2000: data acquisition unit for OxyLite and OxyFlo instruments (Oxford Optronix, Oxford, U.K.)
Equipment available and used in other institutions:
flow cytometer (FACSort Becton Dickinson, Mountain View, CA) Medical faculty, University of Ljubljana
Confocal microscope (Leica TCS SP5, Germany)

20. Institute of Oncology, Ljubljana
Collaborating Institutions
SLOVENIAN:
Institute of Oncology, Ljubljana
Institute of Rehabilitation, Ljubljana
Medical Faculty, University of Ljubljana
Institute Josef Stefan, Ljubljana
INTERNATIONAL:
Institut Gustave-Roussy (Villejuif, France)
Igea S.r.l. (Modena, Italy)
Universität Bielefeld (Bielefeld, Germany)
Faculty of Electrical and Computer Engineering (Zagreb, Croatia)
Institut de Pharmacologie et de Biologie Structurale (Toulouse, France)
Université catholique de Louvain (Brussels, Belgium)
Orthologic Corp. (Tempe, Arizona, USA)
Sewanee University of the South (Sewanee, Tennessee, USA)
Johns Hopkins University (Baltimore, Maryland, USA)

21. Laboratory for bio cybernetics
Peter, Katja, Marko, Jakob, Tadej, David, Matej K., Tomaž, Matej R., Denis,
Blaž, Damijan, Ivan, Janez, Mojca, Stanislav, Gorazd, Vilko, Selma, Čarli,
Nataša, Maša, Alenka, Barbara (mising from the photo: Anže, JernejaTina, Duša)

22. damijan.miklavcic@fe.uni-lj.si http://lbk.fe.uni-lj.si
Thank you !