Projects / Programmes
Electrofusion of cells in biology, biotechnology and medicine
| Code |
Science |
Field |
Subfield |
| 2.06.07 |
Engineering sciences and technologies |
Systems and cybernetics |
Biomedical technics |
| Code |
Science |
Field |
| T115 |
Technological sciences |
Medical technology |
electrofusion, hybrid cells, hybridoma, dielectrophoresis
Researchers (13)
Organisations (1)
Abstract
Electrofusion of biological cells is an efficient and potentially very useful method, but a widespread recognition of its merits has not yet taken place. Cell fusion plays an important role in production of monoclonal antibodies, in immunotherapy of various diseases, as well as in preparation of DNA vaccines.
In the proposed project our aim will be to optimize the parameters influencing cell electrofusion. We will study the influence of the electrical parameters and the properties of the medium on the yield of fused cells, as well as the role of the temperature and integrity of the tubulin cytoskeleton. We will compare the efficiency of two most widely established methods in electrofusion (delivery of a pulse followed by the establishment of contact between the cells, and vice versa). Based on the acquired experimental results we will design and manufacture new devices for electroporators, electrodes and chambers for efficient cell electrofusion. The manufactured instrumentation will also allow for fusion of cells differing in the optimal parameters of electric field.
The main objective of the project is to test the electrofusion method of monoclonal antibodies production and compare it to the established method of chemical fusion using polyethylene glycol (PEG). Hybridoma cell lines will be produced from human lymphocytes and murine myeloma cells, and selective media will be used to isolate heterohybridoma producing a specific antibody.
Significance for science
Electrofusion of biological cells is an efficient and potentially very useful method, but a widespread recognition of its merits has not yet taken place. Cell fusion plays an important role in production of monoclonal antibodies and in immunotherapy of various deseases for preparation of cell vaccines. Lately it was shown that cell fusion is also a mechanism that mediates cell tisue regeneration at cell transplantation. All biological cells are prone to fusion, however, they differ still in electrical and other parameters needed for optimal cell fusion in still an unpredictable manner.
In this project we studied parameters of the electric field, osmolarity and conductivity of cell fusion medium for different cell types. It is important to konow optimal parameters for electroporation of differernt cell types is especially important when different types of the cells are to be fused.
Knowing the values of the parameters that are optimal for as many as possible different cells is important also for the development of the equipment for electrofusion (electroporator, electrodes and chambers). In this field we developed some of the parts needed, such as graphical user interface in .NET environment that is used to controle new prototype of electroporator and a new generator of bipolar electrical pulses or sinusoidal signals for dielectrophoretic pearl chain formation and thua fusion of cells in contact first protocol.
During the development of electrofusion researchers used many different techniques for cell contact achievement. Perfect method that would be suitable for different applications have not been developed yet. During this project we tested some of the described methods for cell contact achievement. By modification of method for fusion of plated cells we developed a new method for cell contact achievement, e.g. modified adherent method. Using this method we produced heterohybridomas between mouse mieloma cells and human lymphocytes B. This is an important achievement since production of human monoclonal antibodies is still problematic in comparisson to animal monoclonal antibodies.
Process of electrofusion is also not explained on molecular level. During this project we started the experiments of studiing cells on the level of tubulin cytoskeleton. These experiments will help to understand the mechanism of electrofusion and to indentify the unknown parameters that influence electrofusion.
Significance for the country
The thematic of electrofusion was incorporated in the International scientific workshop and postgraduate course that was organized in 2009 by the Laboratory of biocybernetics at the Faculty of electrical engineering for the fourth time. This was the enrichment of the program of this school, which was recognized as valuable school by attendees from the whole world.
TRONTELJ, Katja, UŠAJ, Marko. Cell electrofusion. V: KRAMAR, Peter (ur.), KANDUŠER, Maša (ur.). Workbook of the electroporation based technologies and treatments : international scientific workshop and postgraduate course, November 15-21, 2009, Ljubljana, Slovenia. 1. izd. Ljubljana: Založba FE in FRI, 2009, str. 33-35.
Electrofusion is a new field of research and was also transferred in the educational process at the Faculty of electrical engineering.
Most important scientific results
Annual report
2008,
final report,
complete report on dLib.si
Most important socioeconomically and culturally relevant results
Annual report
2008,
final report,
complete report on dLib.si
