Download Electroporation based Technologies andTreatments

Electroporation based
Technologies and Treatments
Damijan Miklavčič
University of Ljubljana
Faculty of Electrical Engineering
Ljubljana, SLOVENIA
“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
Electroporation based Technologies
and Treatments
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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
Electrochemotherapy =
= chemotherapy + electroporation
Electric pulse
generator
Chemotherapeutic
surrounds the cells
Increased membrane Pores reseal
permeability allows
access to the cytosol
Electrodes
time
Tumour
Electric pulse application
Injecting chemotherapeutic
Skin metastasis of squamous cell
carcinoma of the supraglottis
Skin metastasis of malignant
melanoma
Before treatment
One year after ECT
EU 5th FP: Quality of Life
Cliniporator QLK3-1999-00484 (20002003)
– Electro-medical device for ECT
ESOPE QLK3- 2002-02003 (2003-2004)
– European Standard Operating Procedures for
ECT
Objective Response Rate: 85%
No Response:
15%
Final outcome of ESOPE study
90
80
73,7%
70
171 NODULES
%
60
50
40
30
20
11,1%
10,5%
4,7%
10
0
CR
PR
NC
PD
GeneTherapy of diseases with altered angiogenesis:
Psoriasis and Malignant Melanoma
Electroporation of cells:
Experiments and modelling for better understanding
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E
A
B
C
B
A
C
B
100 ms
E
C
1500 ms
3 min
Cell (electro)fusion
• Monoclonal antibody
production
• Cancer immunotherapy
• Receptor insertion into
membrane
• Tissue regeneration
• Research (dissease
models)
Dielectrophoresis
V homogenem električnem polju ni dielektroforeze
Pozitivna dielektroforeza
Negativna dielektroforeza
Biochip and results of experiments
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CHO cellls
Medium conducivity 15mS/cm
negativve dielectrofporesis
frequency 500 Hz
amplitude 4 V
2(1  ) m  (1  2 ) i
3
 p  m
,   1  d / R 
(2  ) m  (1  ) i
Electroporation based Technologies and Treatments
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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
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)
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.
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)
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)
SLOVENIAN:
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Collaborating Institutions
Institute of Oncology, Ljubljana
Institute of Rehabilitation, Ljubljana
Medical Faculty, University of Ljubljana
Institute Josef Stefan, Ljubljana
INTERNATIONAL:
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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)
Laboratory for bio cybernetics
http://lbk.fe.uni-lj.si
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)
Thank you !
[email protected]
http://lbk.fe.uni-lj.si