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Neuronal differentiation of mouse embryonic stem cells using microfluidic spatial confinement of cell parts and soluble factors

Embryonic stem cells provide a potential promising therapeutic approach for tissue regeneration. With the advent of microfluidic technology we can more closely recapitulate features of the native tissue environment. Here we use a contemporary compartmentalized device to culture mouse embryonic stem cells (mESCs) and differentiate them with a defined neuronal medium into process-extending “neurons” that traverse a set of microgrooves from one compartment containing cell bodies to another that is devoid of cell bodies. This device allows spatial isolation of medium and cell parts, and contains a series of microgrooves that guide the paths of neuronal processes. We will use this new culturing approach to determine how various soluble factors or tissue targets might influence neuronal differentiation during development.