摘要
In this talk, we present a microfluidic chip platform integrated with real-time pH sensing, designed to evaluate the therapeutic effects of CAR-NK cells in a hepatocellular carcinoma (HCC) spheroid model. Given that the acidic tumor microenvironment (TME) often impairs immune and drug responses, we developed a pH-sensitive chip based on an Extended-Gate Field-Effect Transistor (EGFET) architecture, integrated into a multilayer PDMS microfluidic system. The pH sensor, utilizing indium tin oxide (ITO) electrodes, demonstrated stable performance over the pH range of 2–8 with a sensitivity of 39.3 mV/pH. Cell experiments involved liver-cell spheroids cultured alone or co-cultured with LX-2 fibroblast-like cells, treated under varying initial pH conditions (pH 4, 6, 8) with CAR-NK, Sorafenib, or their combination. CAR-NK cytotoxicity decreased under acidic conditions, whereas Sorafenib was most effective at a pH of approximately 6. The combination therapy showed enhanced efficacy across all pH levels. Notably, co-cultured Huh7+LX-2 spheroids showed reduced sensitivity to pH changes, suggesting that stromal support buffers metabolic stress and alters treatment response. This platform enables real-time monitoring of the microenvironment and therapeutic evaluation, providing a valuable tool for preclinical immunotherapy research and the development of personalized medicine. To observe the morphology and fluorescence characteristics of hepatocellular carcinoma spheroids under different treatments and pH conditions, this study captured fluorescence microscopy images covering three cell conditions (Lx-2, co-culture, and Huh7) at three pH conditions (pH 4, 6, and 8) to compare the effects of sorafenib treatment on cell morphology. I-Huang Kung 1 and Cheng-Hsien Liu 1, 2, 3 * 1 Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan, ROC 2 Institute of NanoEngineering and MicroSystems, National Tsing Hua University, Hsinchu, Taiwan, ROC 3 College of Semiconductor Research, National Tsing Hua University, Hsinchu, Taiwan, ROC Figure 1