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

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Presentation on theme: "Electroporation based Technologies and Treatments Damijan Miklavčič University of Ljubljana Faculty of Electrical Engineering Ljubljana, SLOVENIA."— Presentation transcript:

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

2 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

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

6 Skin metastasis of squamous cell carcinoma of the supraglottis

7 Skin metastasis of malignant melanoma Before treatmentOne year after ECT

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

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

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 Pozitivna dielektroforeza Negativna dielektroforeza V homogenem električnem polju ni dielektroforeze

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 Gene expression silencing by siRNA Transdermal drug delivery Protein insertion into cell membrane Cell fusion Biotechnology Water and liquid food sterilisation Tissue ablation Electroporation based Technologies and Treatments

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 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) List of other equipment

20 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) Collaborating Institutions

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 Thank you !


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