A microfluidic platform for anterior-posterior human endoderm patterning via countervailing morphogen gradients in vitro

Understanding how morphogen gradients spatially pattern tissues is a fundamental question in developmental biology but can be difficult to directly address using conventional approaches. Here we expose hPSC-derived endoderm cells to countervailing gradients of anteriorizing and posteriorizing signals using a widely available microfluidic device. This approach yielded spatially patterned cultures comprising anterior foregut (precursor to the thyroid, esophagus, and lungs) and mid/hindgut (precursor to the intestines) cells, whose identities were confirmed using single-cell RNA-sequencing. By exposing stem cells to externally applied signaling gradients, this widely accessible microfluidic platform should accelerate the production of spatially patterned tissues, complementing internally self-organizing organoids. Applying artificial morphogen gradients in vitro may also illuminate signaling gradient interpretation mechanisms that are otherwise challenging to study in mammalian embryos in vivo. Overall design: Following differentiation in a microfluidic chip, human pluripotent stem cell-derived anterior and posterior endoderm populations were dissociated and analyzed using single-cell RNA-sequencing.