Literature DB >> 14990328

Setting optimal parameters for in vitro electrotransfection of B16F1, SA1, LPB, SCK, L929 and CHO cells using predefined exponentially decaying electric pulses.

Urska Cegovnik1, Srdjan Novaković.   

Abstract

To achieve the maximal introduction of plasmid DNA into cells and, at the same time, to prevent undesirable cell deaths, electrotransfection conditions should be determined for every single cell type individually. In the present study, we determined the optimal electrotransfection parameters for in vitro transfection of B16F1, SA1, LPB, SCK, L929 and CHO cells. Some of these varying parameters were electric field strength, number of applied pulses and their duration, osmolarity of electroporation buffer, plasmid DNA concentration and temperature at which the electroporation was carried out. The maximal transfection rates at optimal electrotransfection parameters in B16F1, SA1, LPB, SCK, L929 and CHO were 85%, 40%, 60%, 1%, 40% and 65%, respectively. The obtained results confirmed that the electroporation is a useful procedure for an in vitro transfection of the majority of mammalian cells. The method, if optimized, may generate reproducibly high proportion of transfected cells among the cell types that are sensitive to electric field action. Thus, the determined parameters could serve for the subsequent implementations of this method.

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Year:  2004        PMID: 14990328     DOI: 10.1016/j.bioelechem.2003.10.009

Source DB:  PubMed          Journal:  Bioelectrochemistry        ISSN: 1567-5394            Impact factor:   5.373


  8 in total

1.  Analysis and comparison of electrical pulse parameters for gene electrotransfer of two different cell lines.

Authors:  Igor Marjanovic; Sasa Haberl; Damijan Miklavcic; Masa Kanduser; Mojca Pavlin
Journal:  J Membr Biol       Date:  2010-07-20       Impact factor: 1.843

2.  In situ bipolar electroporation for localized cell loading with reporter dyes and investigating gap junctional coupling.

Authors:  Elke De Vuyst; Marijke De Bock; Elke Decrock; Marijke Van Moorhem; Christian Naus; Cyriel Mabilde; Luc Leybaert
Journal:  Biophys J       Date:  2007-09-14       Impact factor: 4.033

3.  Transient gene expression in CHO cells monitored with automated flow cytometry.

Authors:  Greg Sitton; Ann Hansgate; Friedrich Srienc
Journal:  Cytotechnology       Date:  2006-11-22       Impact factor: 2.058

4.  Current Progress in Electrotransfection as a Nonviral Method for Gene Delivery.

Authors:  Lisa D Cervia; Fan Yuan
Journal:  Mol Pharm       Date:  2018-06-20       Impact factor: 4.939

5.  Cell-cell electrofusion: optimization of electric field amplitude and hypotonic treatment for mouse melanoma (B16-F1) and Chinese Hamster ovary (CHO) cells.

Authors:  Marko Usaj; Katja Trontelj; Damijan Miklavcic; Masa Kanduser
Journal:  J Membr Biol       Date:  2010-07-14       Impact factor: 1.843

6.  Optimization of electrotransfection conditions of mammalian cells with different biological features.

Authors:  Huichen Guo; Rongzeng Hao; Yanquan Wei; Dehui Sun; Shiqi Sun; Zhencang Zhang
Journal:  J Membr Biol       Date:  2012-07-27       Impact factor: 1.843

7.  Surviving high-intensity field pulses: strategies for improving robustness and performance of electrotransfection and electrofusion.

Authors:  V L Sukhorukov; R Reuss; D Zimmermann; C Held; K J Müller; M Kiesel; P Gessner; A Steinbach; W A Schenk; E Bamberg; U Zimmermann
Journal:  J Membr Biol       Date:  2005-08       Impact factor: 1.843

8.  Transfection by Electroporation of Cancer and Primary Cells Using Nanosecond and Microsecond Electric Fields.

Authors:  Eivina Radzevičiūtė; Veronika Malyško-Ptašinskė; Jurij Novickij; Vitalij Novickij; Irutė Girkontaitė
Journal:  Pharmaceutics       Date:  2022-06-11       Impact factor: 6.525

  8 in total

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