Preliminary Study Of The Microelectrode Array Based Heterogenous Cell Electrofusion And A Fusion Signal Generator

Cell electrofusion is the technology in which electric methods are used to let two or more cells (protoplasts) contact and then fuse their membrane, cytoplasm and nuclear. The hybrid obtains genetic materials from two or more parental cells, and it can be cultivated to cell engineering products. This method has important significance in the preparation of monoclonal antibody and antitumor vaccine, breeding of animal and microorganism, gene locating, and genetic map drawing. Cell electrofusion has many outstanding advantages such as high precise and controllable operation, wide application, high fusion efficiency, and low cell consumption. However, plate electrode is used in the traditional electrofusion method. Long distance between two electrodes requires high electric voltage. Thus, the generator is required to generate high-voltage signal, and it is difficult for manufacturing and expensive.On the microelectrode array based microfluidic cell-electrofusion chip, a great number of microelectrode pairs are integrated to achieve high-throughput and high-efficiency fusion. Transparent chip is benefit for the manipulation and observation. Futhermore, short distance between two microelectrodes may decrease required the voltage and the difficulty in generator manufacturing. Previous research was focused on the design and fabrication of chip and fusion of homologous cells. In order to validate its practicability, we have carried out some exploratory research on the electrofusion of heterogenous cells. We have analyzed the alignment and fusion of heterogenous cells, and then labeled these cells for clear observation. The alignment and fusion efficiency are also computed, and the fused cells are screened and cultivated. The experimental results show that low driven voltage can be used for electrofusion. Fused cells can be cultivated and grow. Its efficiency is much higher than traditional method. This study is an important reference for other similar research and the design of a low-voltage (as low as tens of volts) signal generator. On the chip, cells are aligned by the dielectrophoresis force. Then, electric pulses are used for reversible perforation. Finally, cytoplasm is exchanged between linked cells. According to the required signals and the range of parameters from above working, the preliminary study of the signal generation is carried out and a new signal generating electric circuit is designed, which can generate mono-frequent sinusoidal signal and low-voltage pulse signal. It set up an important base for the construction of a whole microchip based cell-electrofusion system.