A low-cost multi-layer microfluidic platform for on-site viability assessment of stem cells at single-cell resolution

Abstract

The detection of stem cell activity is crucial for regenerative medicine and cell therapy. However, conventional methods for assessing cell viability are often time-consuming, costly, and result in the non-reusability of cells. In response to these challenges, we developed a low cost and label-free method for detection of cell viability with single-cell resolution, which is implemented by an integrated multi-layer microfluidic system (MLMS) enabling both cell culture and impedance sensing in real time. MLMS integrates a live cell-specific adhesion mechanism with single-cell microfluidic impedance detection. By the selective capture of living cells by fibronectin, live and dead cells can be counted separately, thereby allowing for accurate viability assessment. Fluorescent polystyrene microbeads were counted using both image recognition and impedance detection, demonstrating the high counting accuracy of MLMS. The detection accuracy of chips with micropores of various diameters was assessed using polystyrene (PS) beads of varying sizes. The results show that the chip with 60 μm micropores exhibits superior resolution and enables comprehensive cell detection. Furthermore, mixtures containing different ratios of live and dead cells were analyzed to assess cell viability using the MLMS. The obtained results were very close to the expected values. This work introduces a innovative low-cost, label-free, single-cell resolution solution for cell viability detection. Thereby, it hold promising application prospects in the monitoring and control of cell morphology and quantity during the stem cell culture process.