Optimization of a microfluidic tissue dissociation platform with incorporation of an acoustic cell elution module

Single cell analysis studies have dramatically increased in the past decade, transforming our understanding of cellular heterogeneity in healthy and diseased tissues. However, the dissociation of tissues as the first step in single cell analysis procedure continues to introduce artifacts to the result. Conventional dissociation methods include manual processing steps that make this process time consuming, laborious, and erroneous. Cell aggregation causes underestimation of single cell concentration. Moreover, chemical, and enzymatic treatments can damage cellular protein expression in long exposure times. Therefore, it is crucial to develop a universal approach for single cell preparation with minimum processing time. Microfluidic devices propose various options to make single cell preparation, hands-free, accurate, and predictable. Our lab has developed a microfluidic platform including Digestion, Dissociation, and Filtration devices for tissue dissociation to single cells to automate the single cell preparation while optimizing single cell yields and maintaining cell viability. Each device has its standalone function whereas it can be integrated with other devices in the platform. I optimize the processing conditions for the dissociation of heat and liver tissue as two soft and rigid tissues to achieve highest viability and cell yield. In the next step, dissociation of aggregates with the Integrated Dissociation/Filtration (IDF) device is studied to understand the involved dissociation mechanisms of each module and their connection with cell-cell and cell-ECM interactions to improve cell yield for each cell type. This study will help us to optimize processing condition toward higher cell yield while reducing the processing time. Then, the effect of pulsatile flow versus the steady shear flow is studied on performance of IDF in tissue dissociation. Next, Digestion device is added before IDF device to study the effect of flow rate on cell liberation rate. Finally, a new device will be developed to be integrated with this platform. This device will continuously separate cells from our platform with acoustic forces to increase the overall efficiency in terms of cell yield and functionality