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Electromanipulation of mammalian cells: fundamentals and application

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6 Author(s)
Zimmermann, Ulrich ; Lehrstuhl fur Biotechnol., Wurzburg Univ., Germany ; Friedrich, U. ; Mussauer, H. ; Gessner, P.
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Electroinjection of membrane-impermeable xenomolecules into freely suspended mammalian cells (so-called electroporation) and cell-to-cell electrofusion are powerful tools for manipulation of the genom and the cytosol of cells. Both field pulse techniques are based on the temporary increase of the membrane permeability due to reversible electrical breakdown of the plasma membrane upon application of external high-intensity field pulses of very short duration. Membrane charging and permabilization caused by high-intensity field pulses are preceded and accompanied by transient electrodeformation forces, which lead to an elongation of the cells in low-conductivity media, thus affecting the membrane area of electropermabilization in response to a breakdown pulse. Transient stretching force assumes a maximum value in low-conductivities pulse media. This facilitates incorporation of membrane-impermeable xenomolecules and field-mediated hybridization as well. Therefore, high and reproducible fields of (genetically) manipulated cells can be expected provided that: 1) the duration of the high-intensity field pulses does not exceed shout 100 μs and 2) that the (pulse or fusion) media are hypo-osmolar and exhibit a relatively low conductivities. Such media are also beneficial because field-inducted apoptosis does not occur under these conditions (in contrast to highly conductive media). Indeed, electroporation and electrofusion protocols that fulfill these requirements lead: 1) to high incorporation rates of plasmids (DNA) or artificial chromesomes into living cells without deterioration and 2) to the production of hybridoma cells (by fusion of tumor-infiltrating lymphocytes with heteromyeloma cells), which secrete functional human monoclonal antibodies. Human monoclonal antibodies that bind to and induce apoptosis in autologous tumor cells are promising gents for cancer treatment, as shown by first clinical trials

Published in:

Plasma Science, IEEE Transactions on  (Volume:28 ,  Issue: 1 )

Date of Publication:

Feb 2000

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