Transcriptomically Guided Mesendoderm Induction of Human Pluripotent Stem Cells Using a Systematically Defined Culture Scheme

Abstract

Summary: Human pluripotent stem cells (hPSCs) are an essential cell source in tissue engineering, studies of development, and disease modeling. Efficient, broadly amenable protocols for rapid lineage induction of hPSCs are of great interest in the stem cell biology field. We describe a simple, robust method for differentiation of hPSCs into mesendoderm in defined conditions utilizing single-cell seeding (SCS) and BMP4 and Activin A (BA) treatment. BA treatment was readily incorporated into existing protocols for chondrogenic and endothelial progenitor cell differentiation, while fine-tuning of BA conditions facilitated definitive endoderm commitment. After prolonged differentiation in vitro or in vivo, BA pretreatment resulted in higher mesoderm and endoderm levels at the expense of ectoderm formation. These data demonstrate that SCS with BA treatment is a powerful method for induction of mesendoderm that can be adapted for use in mesoderm and endoderm differentiation. : In this article, Carpenedo and Stanford and colleagues demonstrate a robust and reproducible single-cell seeding method for rapid induction of mesendoderm for hPSCs. Transcriptomic data indicated that the method could be applied to mesoderm and endoderm differentiation protocols, which was demonstrated experimentally. Formation of mesoderm and endoderm following pre-differentiation was further demonstrated in long-term in vitro and in vivo assays. Keywords: mesendoderm, human pluripotent stem cells, defined differentiation, human embryonic stem cells, human induced pluripotent stem cells