Research On A Portable Constant Temperature Perfusion Cell Culture System Based On Microfluidic Chips

Cell is the basic unit of life activity. and it is also the basis of the study of various life phenomena. Observing the growth of cells, for exploring the law of life and disease prevention, diagnosis and treatment, is of great significance. Due to its miniaturization and high integration, and other characteristics, as well as the advantages of fast and efficient, A microfluidic chip can realize the control of some micron size of the objects by integrating other functional components, such as particle, cells,etc. Compared with the traditional method in the study of single cell and high throughput, the microfluidic chip has the advantages of high ratio of performance to price ratio and low sample consumption, and so on i t has broad application prospects in the field of life science and chemistry.In this article, with comparing and analyzing the advantages and disadvantages of conventional cell culture method and microfludic cell culture system, a constant temperature cell culture system has been designed and fabricated based on micro fluidic technology. The experiment of perfusion cell culture has also been carried out to verify the function of the system. The Film thermistor temperature sensor and PID intelligent temperature controller are selected as the constant temperature control system. To explore the constant temperature control conditions that can satisfy the normal growth of cells, Multiphysics COMSOL 4.4 is used to simulate the temperature field for cell culture microfluidic chip and thermostatic culture chamber. ITO conductive glass is used to produce a thermostat heater by the process steps of cleaning, etching, electrode forming and so on. A programmable single axis stepper motor, driver and controller are selected to build the cell culture fluid circulation system. Using the cured PDMS as the top material, the microfluidic chip with poly methyl methacrylate as material is processed by numerical control milling machine, and with high frequency plasma glow discharge apparatus to combine the two. In this article, the constant temperature cell culture system is divided into three parts which are assembled: the cell culture microfluidic chip, the cell culture fluid circulation system, and the cell culture box with constant temperature. When the temperature is set to 310.15 ±0.2K, the stability test is carried out, thus confirmed the feasibility of this system.On the microfluidic cell culture platform,a control experiment has been conducted on a continuous 96 h of yeast. After comparison and analysis of the experimental results,it has been confirmed that the constant and suitable culture temperature and the confined culture environment have important function and meaning to cell culture. The constant temperature cell culture system is verified to have the characteristics of high efficiency,superiority and practicability in cell culture. The critical frequency of the yeast cells has been measured by using the micro electrode, and the reference will be provided for the subsequent on-chip cell culture and the related experiments.