一种用于捕获口腔鳞癌细胞微流控芯片系统的开发和初步应用OACSTPCD

Objective:The aim of this study was to investi-gate the feasibility of capturing oral squamous carcinoma cells by microfluidic chip using biophilic method,and to provide theoretical basis for the development of microfluidic chip for early diagnosis and screening of oral squamous carcinoma.Methods:Firstly,the plasmid with mCherry fluorescence was trans-fected into human tongue squamous carcinoma cells(Cal 27)using lentiviral transfection method for subsequent more intui-tive observation in microfluidic chip.Next,COMSOL Multiphysics software was used to simulate the liquid flow and cell movement inside the microfluidic chip to optimize the shape and structure of the chip,designing a suitable chamber shape.And a model was designed to obtain a silicon mold by ordinary photolithography,which was inverted,bonded.The anti-EpCAM antibody was ligated in the chamber using chemical bonds,realizing the cell capture function of the microfluidic chip.Finally,the effects on the capture efficiency of the microfluidic chip were investigated on the variables of inlet flow rate,chamber height,and antibody concentration,and the optimal experimental parameters were screened.Results:Lentivirus suc-cessfully transfected mCherry fluorescence into Cal 27 cells with an average fluorescence expression rate of 95.6％.The mi-crofluidic chip with microcolumn structure fabricated had a capture efficiency of up to 70％.It was also concluded that the capture efficiency was improved under the conditions of 10 μL/min(P＜0.001)inlet flow rate and 8 μg/mL(P＜0.01)antibody concentration,while the difference in chamber height did not affect the capture efficiency(P＞0.05).Conclusion:Microfluidic chip designed in this study can capture human tongue squamous carcinoma cells(Cal 27).It not only proves that the use of microfluidic chip can capture and separate oral squamous carcinoma cells,but also provides an important theoreti-cal basis for the development of microfluidic chip for early screening of oral squamous carcinoma.