A PROCEDURE OF EVALUATING THE YIELD OF HYBRIDOMA CELLS AND CELL PRODUCTS BY USING CONFOCAL MICROSCOPY AND AN AUTOCLAVABLE ELECTROFUSION CHAMBER

摘要

The subject of the invention in the field of biomedical technology is the evaluation procedure for the determination of the yield of hybrid cells - hybridomas in a cell vaccine and cell products, which consists of measurements of the area of co-localized pixels of an image recorded by multichannel confocal microscopy and by using a reusable autoclavable electrofusion chamber to be used in the preparation of hybrid cells. The quantification of the yield of hybrid cells is based on measurements of the area of co-localized pixels and allows simultaneous visualization and documentation of the procedure. This approach has a wider applicability, for example also in cases in which a fraction of one or multiple cell types are present in a suspension or a structure. The subject of this invention is also a corresponding autoclavable electrofusion chamber, more precisely, a chamber for fusion of biological cells in electric field. The invention brings a new construction of a re-usable electrofusion chamber. The chamber consists of a top part (EI.A), a bottom part (EI.B), a glass plate that fits in between the two parts of the chamber and carries electrodes, an O-ring, a cover and nuts. All parts are autoclavable separately and can be easily assembled into a functional sterile fusion chamber in a sterile hood area. In addition to this, by exchanging one part of the electrofusion chamber it is possible to change the area of the electrofusion surface and therefore expose different volumes of cells to the electrofusion process. The electrofusion chamber is made from biocompatible materials and is suitable for the preparation of hybrid cells for vaccines to be used in human therapy.