Simulation And Experimental Study On Silicon-glass Based Cell Electro-fusion Chip

Cell fusion technology is also called cell hybridization, in which two or more cells (protoplasts) are contacted one by one and then their membrane, cytoplasm and nuclear are fused by using some biological, chemical or physical methods. The hybrid may obtains genetic material from two or more parent cells. Thus, it achieves new biological or genetic characteristics and may be used to develop new species, strain or cell engineering products. The development of cell fusion technology has gone through three stages: biological, chemical and physical inducement, in these cell electrofusion was developed as a new physical mediated way for cell fusion. Compared to traditional cell fusion methods, cell electrofusion technology has many advantages such as easy operation, high efficiency, harmless to cells, suitable for instrumentation applications, and so on. However, there are still some problems in the traditional cell electrofusion method including difficult to observing and high voltage required, which hindered its wider use. In order to solve these problems, a new cell electrofusion technology based on chip has been set up. In our group, related study has also been carried out and several kinds of cell electrofusion chips has been developed, which could be used for cell fusion under low-voltage driven. According to the previous studies, a new chip based on silicon-glass substrates is developed in our group in order to further improve the chip architecture and achieve better fusion effect. This chip can not only reduce the drive voltage and improve fusion efficiency, but also can be used under an inverted fluorescence microscope to observe the fusion of fluorescence labeled cells.In this thesis, first the simulation of the electric field within both silicon-glass based chip and SOI based chip is be done. The simulation results show that using the silicon-glass based chip can greatly reduce driven voltage, and the same size of the micro-channel and electrodes structure design makes the electric field distribute more uniform, which can effectively improve the cell fusion efficiency. The simulation results provide a strong theoretical support to chip fabrication and packaging. Then the packaged chip is used to do some alignment and fusion experiments on HEK-293 and NIH-3T3 cells. Experimental and simulation results are matched each other, using the driven voltage under 10V could induce cell alignment and fusion, which is much lower than using previous chips, and the uniform electrodes structure also improves the fusion efficiency. Otherwise, the experiment on heterologous cell alignment and fusion of HEK-293 and NIH-3T3 is done. Cell-fusion process of different kinds of fluorescent cells is directly observed under the fluorescence microscope, and survival heterozygous cells are obtained, which validates the advantage of this new technology and lay foundation for the further fusion research of somatic cells and stem cells.