Laboratory excercise is designed so that it enables participants to do the experiment of electrofusion by means of modified adherence method. Cells, dyed with fluorescent dyes CMRA and CMFDA, are plated and allowed to adhere slightly to the surface of the plate. Cells are then exposed to hypoosmolar buffer and electroporated after appropriate time. The results are evaluated on a fluorescent microscope.
F.18 Transfer of new know-how to direct users (seminars, fora, conferences)COBISS.SI-ID: 7425108
The developed and protected electrode tip chamber is an important improvement of the electroporation, electrofusion and electrotransfection processes, since it enables electropermeabilization of cells in an uniform electric field with reduced number of cellmanipulations needed.
F.33 Slovenian patentCOBISS.SI-ID: 6613076
Modified adherence method was described for the first time. This method uses cells that was allowed to attach slightly to the surface and retain sferical shape instead of using the confluent cell culture. The described method for cell contact achievement was used on two cell lines. The results show that fusion yield and viability of cells after electroporation are cell line dependent. Yields of fused cells, obtained with the described method was comparable with results of other published researches.
B.03 Paper at an international scientific conferenceCOBISS.SI-ID: 7081812
Pipette tip with integrated electrodes, developed in in our laboratory, was tested for electroporation and electrotransfection of CHO cells. In this work we showed that pipette tip with integrated electrodes is suitable equipment for electroporation of cells in suspension and for transfection of cells. Electric pulses in different directions were used that were more succesful at electroporation of cells than pulses in one direction.
B.03 Paper at an international scientific conferenceCOBISS.SI-ID: 7081556
It is known that hypotonic fusion buffer improves electrofusion efficiency. We determined that different cell lines differ in their tolerance for incubation in hipoosmolar conditions. Swelling dynamics must be therefore determined for every cell line. The duration of increased cell volume is prolonged by electroporation and can improve electrofusion efficiency. Viability of cells after incubation in hypotonic buffer was not affected.For determination of electroporation parameters for electroporation of cells in hypotonic buffer, swelling of the cells must be taken into account.
B.03 Paper at an international scientific conferenceCOBISS.SI-ID: 6436692