Defined human pluripotent stem cell culture enables highly efficient neuroepithelium derivation without small molecule inhibitors.

摘要: The embryonic neuroepithelium gives rise to the entire central nervous system in vivo, making it an important tissue for developmental studies and a prospective cell source regenerative applications. Current protocols deriving homogenous neuroepithelial cultures from human pluripotent stem cells (hPSCs) consist of either embryoid body-mediated neuralization followed by manual isolation step or adherent differentiation using small molecule inhibitors. Here, we report that hPSCs maintained under chemically defined, feeder-independent, xeno-free conditions can be directly differentiated into pure ([mt]90% Pax6(+)/N-cadherin(+) with widespread rosette formation) within 6 days conditions, without inhibitors, only minimalistic medium consisting Dulbecco's modified Eagle's medium/F-12, sodium bicarbonate, selenium, ascorbic acid, transferrin, insulin (i.e., E6 medium). Furthermore, provide evidence defined culture enable this high level neural conversion contrast on mouse fibroblasts (MEFs). In addition, previously MEFs could rapidly converted compliant state upon transfer these while still maintaining their ability generate all three germ layers. Overall, fully scalable protocol should broadly useful generating therapeutic