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Stem Cell Reports. 2019 Sep 10;13(3):499-514. doi: 10.1016/j.stemcr.2019.07.014. Epub 2019 Aug 15.

Generation of a Purified iPSC-Derived Smooth Muscle-like Population for Cell Sheet Engineering.

Author information

1
Center for Regenerative Medicine, Boston University and Boston Medical Center, 670 Albany Street, 2(nd) Floor, Boston, MA 02118, USA; Department of Biomedical Engineering, Boston University, 44 Cummington Mall, Boston, MA 02215, USA.
2
Center for Regenerative Medicine, Boston University and Boston Medical Center, 670 Albany Street, 2(nd) Floor, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, 72 East Concord Street, Boston, MA 02118, USA.
3
Department of Biomedical Engineering, Boston University, 44 Cummington Mall, Boston, MA 02215, USA. Electronic address: jywong@bu.edu.
4
Center for Regenerative Medicine, Boston University and Boston Medical Center, 670 Albany Street, 2(nd) Floor, Boston, MA 02118, USA; The Pulmonary Center and Department of Medicine, Boston University School of Medicine, 72 East Concord Street, Boston, MA 02118, USA. Electronic address: dkotton@bu.edu.

Abstract

Induced pluripotent stem cells (iPSCs) provide a potential source for the derivation of smooth muscle cells (SMCs); however, current approaches are limited by the production of heterogeneous cell types and a paucity of tools or markers for tracking and purifying candidate SMCs. Here, we develop murine and human iPSC lines carrying fluorochrome reporters (Acta2hrGFP and ACTA2eGFP, respectively) that identify Acta2+/ACTA2+ cells as they emerge in vitro in real time during iPSC-directed differentiation. We find that Acta2hrGFP+ and ACTA2eGFP+ cells can be sorted to purity and are enriched in markers characteristic of an immature or synthetic SMC. We characterize the resulting GFP+ populations through global transcriptomic profiling and functional studies, including the capacity to form engineered cell sheets. We conclude that these reporter lines allow for generation of sortable, live iPSC-derived Acta2+/ACTA2+ cells highly enriched in smooth muscle lineages for basic developmental studies, tissue engineering, or future clinical regenerative applications.

KEYWORDS:

cell sheets; directed differentiation; fluorochrome reporters; induced pluripotent stem cells; smooth muscle cells

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