Agarose microgel culture delineates lumenogenesis in naive and primed human pluripotent stem cells

• Published in: Stem cell reports 2021-05, Vol.16 (5), p.1347-1362
• Main Authors: Schindler, Magdalena, Siriwardena, Dylan, Kohler, Timo N., Ellermann, Anna L., Slatery, Erin, Munger, Clara, Hollfelder, Florian, Boroviak, Thorsten E.
• Format: Article
• Language: English
• Subjects: Cell Culture Techniques; Cell Differentiation - drug effects; Cell Lineage - drug effects; Cell Survival - drug effects; Cells, Immobilized - cytology; Cells, Immobilized - drug effects; epiblast; epiblast spheroid; Germ Layers - cytology; human development; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - metabolism; lumenogenesis; microfluidic platform; microgel suspension culture; Microgels - chemistry; Morphogenesis - drug effects; naive pluripotency; primed pluripotency; Protein Kinase Inhibitors - pharmacology; Resource; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - metabolism; self-organization; Sepharose - pharmacology; Spheroids, Cellular - cytology; Spheroids, Cellular - drug effects
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2021.04.009

Human periimplantation development requires the transformation of the naive pluripotent epiblast into a polarized epithelium. Lumenogenesis plays a critical role in this process, as the epiblast undergoes rosette formation and lumen expansion to form the amniotic cavity. Here, we present a high-throughput in vitro model for epiblast morphogenesis. We established a microfluidic workflow to encapsulate human pluripotent stem cells (hPSCs) into monodisperse agarose microgels. Strikingly, hPSCs self-organized into polarized epiblast spheroids that could be maintained in self-renewing and differentiating conditions. Encapsulated primed hPSCs required Rho-associated kinase inhibition, in contrast to naive hPSCs. We applied microgel suspension culture to examine the lumen-forming capacity of hPSCs and reveal an increase in lumenogenesis during the naive-to-primed transition. Finally, we demonstrate the feasibility of co-encapsulating cell types across different lineages and species. Our work provides a foundation for stem cell-based embryo models to interrogate the critical components of human epiblast self-organization and morphogenesis. [Display omitted] •High-throughput platform to encapsulate human PSCs in agarose microgels•Human PSCs self-organize into epiblast spheroids and undergo lumen expansion•Lumen expansion increases in primed versus naive human PSCs•Monodisperse co-culture of embryonic and extraembryonic stem cells In this article, Boroviak and colleagues present a high-throughput platform to encapsulate human PSCs into agarose microgels. Human PSCs undergo lumenogenesis and self-organize into rosette-like structures, similar to the human peri-implantation embryo. Their microgel suspension culture regime provides a foundation for the controlled assembly of stem cell-based embryo models in our own species.