A rapid and high-throughput microfluidic stem cell analyzer

Current methods to discern the differentiation state of stem cells are time-consuming, labor-intensive, costly, invasive, and consequently, ill-suited for clinical applications. To overcome these limitations, we propose to develop a microfluidic stem cell analyzer for accurate, non-invasive identification of the cell differentiation state in conjunction with automated cell sorting at high-throughput. Our technology enables significant improvements in analysis speed, non-invasiveness, device automation, and marked reduction in logistical burden and operating cost. In Phase I, key technology concepts were successfully demonstrated. Two different designs were developed using physics-based simulations, followed by state-of-the-art fabrication and engineering. Experiments were undertaken to demonstrate microfluidic stem cell manipulation and impedance identification of the cell differentiation state, which firmly established proof-of-concept of the technology. In Phase II, efforts will focus along two directions. First, an advanced cell sorter will be developed for separating cells of different characteristics. Design optimization and experimental characterization will be carried out for enhanced performance. Second, the microfluidic stem cell analyzer will be integrated with COTS component technologies to develop an integrated prototype for automated operation. The functionality of differentiation state identification and cell sorting will be extensively demonstrated using human iPSCs progressing along various lineage commitment pathways. A multi-disciplinary team with experience in all aspects of the proposed effort including microfluidics, stem cell bioengineering, and systems engineering has been assembled to ensure successful completion of project milestones.