Leveraging dynamic cell responses to design new protocols for pluripotency maintenance

They are unspecialized cells that can be indefinitely expanded retaining pluripotency or pushed into specific differentiated states by proper stimuli. Various protocols have been proposed to maintain mouse embryonic stem cells (mESCs) in the pluripotent state; state of the art cell culture protocols are based on serum-free media, and continuous provision of drugs inhibiting pathways associated to differentiation.

We explore here the possibility to avoid continuous drug exposure. First, we employ an external feedback control microfluidics/microscopy platform to steer the expression of a pluripotency reporter. Control experiments show a significant delay in cell response to drugs; these dynamics are confirmed by flow-cytometry open-loop experiments.