Tracking and Predicting Reprogramming from a Biophysical Perspective

Molecular mechanisms of reprogramming terminally differentiated cells to a pluripotent state still are poorly understood. As a result, standard reprogramming techniques are noisy and inefficient. To address these shortcomings, we have developed a novel platform that allows for the dynamic tracking of subpopulations in a longitudinal manner across long periods of time. This platform is a simple approach that combines live-cell microscopy with surface-modified multiwell plates that separates thousands of cell populations. With this we are able to both watch and physically constrain cells into discrete islands during reprogramming. By watching subpopulations in real time we are able to distinguish intermediate states that either contribute or are detrimental to complete reprogramming. Furthermore by controlling the island geometry at the microscale, we are able to selectively activate mechanotransduction pathways (e.g., YAP/Taz) to promote transition through the endothelial-mesenchymal transition during the reprogramming process. Direct manipulation of the YAP/Taz pathway via lentiviral overexpression also affected the reprogramming process. Overall, this simple platform allows us to expand our understanding of intermediate cell states and increasing the overall efficiency of the reprogramming process.