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

Richard L Carpenedo; Sarah Y Kwon; R Matthew Tanner; Julien Yockell-Lelièvre; Chandarong Choey; Carole Doré; Mirabelle Ho; Duncan J Stewart; Theodore J Perkins; William L Stanford

doi:10.1016/j.stemcr.2019.11.001

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

Stem Cell Reports. 2019 Dec 10;13(6):1111-1125. doi: 10.1016/j.stemcr.2019.11.001.

Affiliations

¹ The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Ottawa Institute of Systems Biology, Ottawa, ON, Canada. Electronic address: rcarpenedo@ohri.ca.
² The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
³ The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Ottawa Institute of Systems Biology, Ottawa, ON, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada.
⁴ The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Sinclair Centre for Regenerative Medicine, Ottawa Hospital Research Institute, Ottawa, ON, Canada.
⁵ The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Ottawa Bioinformatics Core Facility, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON, Canada.
⁶ The Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Ottawa, ON, Canada; Ottawa Institute of Systems Biology, Ottawa, ON, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON, Canada; Department of Biochemistry, Microbiology, and Immunology, University of Ottawa, Ottawa, ON, Canada. Electronic address: wstanford@ohri.ca.

Abstract

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.

Keywords: defined differentiation; human embryonic stem cells; human induced pluripotent stem cells; human pluripotent stem cells; mesendoderm.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Activins / pharmacology
Bone Morphogenetic Protein 4 / pharmacology
Cell Culture Techniques
Cell Differentiation / drug effects
Cell Differentiation / genetics*
Cells, Cultured
Endoderm / cytology
Endoderm / metabolism
Gene Expression Profiling
Humans
Mesoderm / cytology*
Mesoderm / metabolism*
Pluripotent Stem Cells / cytology*
Pluripotent Stem Cells / drug effects
Pluripotent Stem Cells / metabolism*
Single-Cell Analysis
Teratoma / etiology
Time Factors
Transcription, Genetic*
Transcriptome

Substances

BMP4 protein, human
Bone Morphogenetic Protein 4
activin A
Activins

Grants and funding

MOP-89910/CIHR/Canada