A microfluidic platform for anterior-posterior human endoderm patterning via countervailing morphogen gradients in vitro
Leeya Engel,
Kevin J. Liu,
Kiara W. Cui,
Eva L. de la Serna,
Vipul T. Vachharajani,
Carolyn E. Dundes,
Sherry Li Zheng,
Manali Begur,
Kyle M. Loh,
Lay Teng Ang,
Alexander R. Dunn
Affiliations
Leeya Engel
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA; Faculty of Mechanical Engineering, Technion – Israel Institute of Technology, Haifa 3200003, Israel
Kevin J. Liu
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA
Kiara W. Cui
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA
Eva L. de la Serna
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA
Vipul T. Vachharajani
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA; Program in Biophysics, Medical Scientist Training Program, Stanford University, Stanford, CA 94305, USA
Carolyn E. Dundes
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA
Sherry Li Zheng
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA
Manali Begur
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA
Kyle M. Loh
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Developmental Biology, Stanford University, Stanford, CA 94305, USA
Lay Teng Ang
Institute for Stem Cell Biology & Regenerative Medicine, Stanford University, Stanford, CA 94305, USA; Department of Urology, Stanford University, Stanford, CA 94305, USA; Corresponding author
Alexander R. Dunn
Department of Chemical Engineering, Stanford University, Stanford, CA 94305, USA; Corresponding author
Summary: 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 (scRNA-seq). 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 how developing tissues interpret signaling gradients in systems that are not readily accessible for in vivo studies.
