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Query: "keywords" (electroporation) .

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Cell size dynamics and viability of cells exposed to hypotonic treatment and electroporation for electrofusion optimization
Marko Ušaj, Katja Trontelj, Rosana Hudej, Maša Kandušer, Damijan Miklavčič, 2009, original scientific article

Abstract: Background. Various electrofusion parameters have to be adjusted to obtain theoptimal electrofusion efficiency. Based on published data, good electrofusion conditions can be achieved with the hypotonic treatment. However, the duration of the hypotonic treatment before electroporation and buffer hypoosmolarity have to be adjusted in order to cause cell swelling, to avoid regulatory volume decrease and to preserve cell viability. The aims of our study were to determine cell size dynamics and viability of four different cell lines in hypotonic buffer and to study the influence of the electroporation on the selected cell line in hypotonic buffer. Materials and methods. Cell size dynamics of different cell lines exposed to hypotonic buffer and electroporation were analyzed by time-resolvedcell size measurements. The viability of hypotonically treated oržand electroporated cells was determined 24 h after the experiment by a modified crystal violet (CV) viability assay. Results. In our experimental conditions the hypotonic treatment at 100 mOsm was efficient for CHO, V79 and B16-F1 cell lines. The optimal duration of the treatment was between two and five minutes. On the other hand the same hypotonic treatment did not cause cell swelling of NS1 cells. Cell swelling was also observed after electroporation of B16-F1 in isotonic buffer and it was amplified when hypotonic buffer was used. In addition, the regulatory volume decrease was successfully inhibited with electroporation. Conclusions. Cell size dynamicsin hypotonic conditions should be studied for each cell line since they differ in their sensitivity to the hypotonic treatment. The inhibition of cell regulatory volume decrease by electroporation may be beneficial in achieving higher electrofusion efficiency. (Abstract truncated at 2000 characters)
Keywords: hypotonic treatment, cell swelling, regulatory volume decrease, cell size measurements, viability, electrofusion, electroporation
Published in DiRROS: 08.03.2024; Views: 111; Downloads: 32
.pdf Full text (263,01 KB)

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Numerical modeling in electroporation-based biomedical applications
Nataša Pavšelj, Damijan Miklavčič, 2008, original scientific article

Abstract: Background. Numerous experiments have to be performed before a biomedical application is put to practical use in clinical environment. As a complementary work to in vitro, in vivo and medical experiments, we can use analytical and numerical models to represent, as realistically as possible, real biological phenomena of, in our case, electroporation. In this way we canevaluate different electrical parameters in advance, such as pulse amplitude, duration, number of pulses, or different electrode geometries. Suchnumerical models can contribute significantly to the understanding of an experiment and treatment planning as well as to the design of new electroporation devices and electrodes. Methods. We used commercially available modeling software, based on finite element method. We constructed a model of a subcutaneous tumor during electrochemotherapy (EMAS) and a model ofskin during gene electrotransfer (COMSOL Multiphysics). Tissue-electrode geometries, pulse parameters and currentvoltage measurements from in vivo experiments were used to develop and validate the models. Results. To describeadequately our in vivo observations, a tissue conductivity increase during electroporation was included in our numerical models. The output currents of the models were compared to the currents and the voltages measuredduring in vivo experiments and a good agreement was obtained. Also, when comparing the voltages needed for a successful electropermeabilization assuggested by the models, to voltages applied in experiments and achieving a successful electrochemotherapy or in vivo gene electrotransfer, good agreementcan be observed. Conclusions. Modeling of electric current and electric field distribution during cell and tissue electroporation proves to be helpful in describing different aspects of the process and allowing us to design electrodes and electroporation protocols as a part of treatment planning.
Keywords: electroporation, gene electrotransfer, electrochemotherapy, subcutaneous tumor, finite-element method
Published in DiRROS: 07.03.2024; Views: 101; Downloads: 31
.pdf Full text (549,62 KB)

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Electroporator for in vitro cell permeabilization
Marko Puc, Karel Flisar, Stanislav Reberšek, Damijan Miklavčič, 2001, original scientific article

Abstract: The use of high voltage electric pulse technology, electroporation, in cell biology, biotechnology and medicine has attracted an enormous interest. Electroporation is a transient phenomenon that increases the permeability of cell plasma membrane. In the state of high permeability, the plasma membrane allows small and large molecules to be introduced into the cytoplasm, althoughthe cell plasma membrane represents a considerable barrier for them inits normal state. The effectiveness of electroporation depends on many parameters that can be divided into the parameters of the electric field and the parameters that define the state of cells and their surrounding i.e, temperature, osmotic pressure, etc. In this article, we present a prototype electroporator GT-1 for in vitro electropermeabilization that we have developed. Our electroporator offers a vast flexibility of parameters and can generate high and low voltage pulses, of which the latter ones are used for electrophoretic transfer of charged molecules through permeabilized cell plasma membrane.
Keywords: electroporation instrumentation methods, cell membrane permeabilization
Published in DiRROS: 25.01.2024; Views: 178; Downloads: 40
.pdf Full text (239,19 KB)

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Requirements for a clinical electrochemotherapy device - electroporator
Marko Puc, Stanislav Reberšek, Damijan Miklavčič, 1997, original scientific article

Keywords: electroporation instrumentation, neoplasms, drug therapy
Published in DiRROS: 16.01.2024; Views: 131; Downloads: 41
.pdf Full text (1,38 MB)

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