Actuation Enhances Patterning in Human Neural Tube Organoids

Tissues achieve their complex spatial organization though physical interactions mediated by mechanical forces. Current strategies to generate in-vitro tissues have largely failed to implement such active, dynamically coordinated mechanical manipulations, relying instead on extracellular matrices which respond to, rather than impose mechanical forces. Here we develop devices which enable the actuation of organoids, and show that active mechanical forces increase growth and lead to enhanced patterning in an organoid model of the neural tube derived from single human pluripotent stem cells (hPSC). We demonstrate that organoid mechanoregulation due to actuation operates in a temporally restricted competence window, and that organoid response to stretch is mediated extracellularly by matrix stiffness and intracellularly by cytoskeleton contractility and planar cell polarity. Exerting active mechanical forces on organoids using the approaches developed here is widely applicable and should enable the generation of more reproducible, programmable organoid shape, identity and patterns, opening avenues for use of these tools in regenerative medicine and disease modelling applications. 5 samples are analyzed to compare the effect of stretching on human neural tube organoids. 2x day 11 samples of stretched and unstretched, 2x day 5 samples of stretched and unstretched (immediately after RA-SAG treatment and stretch completion) and 1 sample at day 3 (immediately before RA-SAG treatment and stretch initiation)