print small

Participating Countries:

Algeria

Argentina

Australia

Austria

Belgium

Bosnia and Herzegovina

Bulgaria

Croatia

Czech Republic

Denmark

Finland

France

FYR of Macedonia

Germany

Greece

Iceland

Ireland

Israel

Italy

Lithuania

Morocco

Netherlands

New Zealand

Poland

Portugal

Romania

Russian Federation

Serbia

Slovenia

Spain

Sweden

Switzerland

Turkey

Ukraine

United Kingdom

United States

Member area provided by LTFE
COST is supported by the EU Framework Programme Horizon 2020
This website is supported by COST
24/10/2012 (Added to site)
Author(s): Usaj, M.; Kanduser, M.

The Systematic Study of the Electroporation and Electrofusion of B16-F1 and CHO Cells in Isotonic and Hypotonic Buffer

Journal: Journal of Membrane Biology, 245 (2012), pp. 583-590
DOI: 10.1007/s00232-012-9470-2
Request reprint  |  Tell your friend  | 

Abstract: The fusogenic state of the cell membrane can be induced by external electric field. When two fusogenic membranes are in close contact, cell fusion takes place. An
appropriate hypotonic treatment of cells before the application of electric pulses significantly improves electrofusion efficiency. How hypotonic treatment improves
electrofusion is still not known in detail. Our results indicate that at given induced transmembrane potential electroporation was not affected by buffer osmolarity. In contrast to electroporation, cells’ response to hypotonic treatment significantly affects their electrofusion. High fusion yield was observed when B16-F1 cells were used;
this cell line in hypotonic buffer resulted in 41 ± 9 % yield, while in isotonic buffer 32 ± 11 % yield was observed. Based on our knowledge, these fusion yields determined in situ by dual-color fluorescence microscopy are among the highest in electrofusion research field. The use of hypotonic buffer was more crucial for electrofusion
of CHO cells; the fusion yield increased from below 1 % in isotonic buffer to 10 ± 4 % in hypotonic buffer. Since the same degree of cell permeabilization was achieved in both
buffers, these results indicate that hypotonic treatment significantly improves fusion yield. The effect could be attributed to improved physical contact of cell membranes
or to enhanced fusogenic state of the cell membrane itself.



Project Office

Working groups

Steering Committee

Founding members

DC Rapporteurs

Related sites: