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Preliminary Study of the Microelectrode Array Based Heterogenous Cell Electrofusion and a Fusion Signal Generator

Author: ZuoJiaWen
Tutor: ZhengXiaoLin
School: Chongqing University
Course: Biomedical Engineering
Keywords: Cell fusion Microfluidic Heterologous cells Dielectrophoretic
CLC: R318.0
Type: Master's thesis
Year: 2011
Downloads: 20
Quote: 0
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Abstract


Cell fusion refers to the in vitro environment by electrical stimulation to promote two or more cells or protoplasts contact with each other and then fused to form a mononuclear or polynuclear hetero nuclear cellular process. The fusion cells have the genetic characteristics of two or more of the parental cells, can develop a new cell engineering products has great significance, especially monoclonal antibodies and anti-tumor vaccine production, animal and plant breeding distant hybridization . Cell fusion technology has a precise and controlled operation, the integration of high efficiency and low cellular demand advantage. However, the conventional electric fusion method using plate electrodes, electrode spacing width, high fusion signal voltage, the signal generating device requires to produce the high-voltage signal, are difficult to produce, the cost is high. Microelectrode array-based microfluidic cell fusion chip, can greatly reduce the drive voltage and the difficulty of making electrical signal generator, to achieve high-throughput cell fusion and fusion efficiency. Research group pre carried out a large number of studies focused on chip design, production and homologous cell fusion. Expand practical the electric fusion experiments in heterologous cells to carry out exploratory research, trying to find a suitable fusion parameters. Queuing and integration, and analysis of heterologous animal cells by the method of fluorescence labeled clear and reliable observation of the effect of the fusion experiment, the researchers. Calculated on this basis, the the queuing rate and the fusion rate of the heterologous cells, Meanwhile, the identification and screening of the fusion cell karyotype. Experimental results show that heterologous cells electrically driven by low voltage fusion is achievable after fusion cells can continue the culture and growth, the fusion efficiency is considerably higher than the conventional method. The parameters of the study can be used as a low-pressure electric fusion important reference control signal generator, electric fusion to the signal voltage from the traditional several hundred volts to tens of volts. Preliminary studies showed that the integration process on the chip power cells by dielectrophoretic force acting on the chip queue; then unipolar narrow pulse, the cells produce a reversible electroporation; last in the dielectrophoretic force the the role of perforation cells further material exchange, complete electric fusion process. Control signal for this process, as well as the range of parameters in the actual study, we carried out a preliminary study of the signal generator cell fusion based on the micro-electrode array, designed and can output 1 MHz, 10 V adjustable sinusoidal signal and the pulse width of 50μs, the number 3, the pulse interval of 1 s, low-voltage electrical pulse signal circuit system construction laid the foundation for the a complete microelectrode array cell electric fusion.

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