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

Autor: Hector A. Marquez, Joyce Wong, Chian Yang, George Kwong, Darrell N. Kotton
Rok vydání: 2019
Předmět:
0301 basic medicine
Cell type
induced pluripotent stem cells
fluorochrome reporters
Myocytes
Smooth Muscle
Population
Gene Expression
Mice
Transgenic
Biology
Biochemistry
Article
Green fluorescent protein
Transcriptome
Mice
03 medical and health sciences
0302 clinical medicine
Directed differentiation
Tissue engineering
Genetics
Animals
Humans
Cell Lineage
Induced pluripotent stem cell
education
lcsh:QH301-705.5
Aorta
Principal Component Analysis
lcsh:R5-920
education.field_of_study
Myosin Heavy Chains
Tissue Engineering
directed differentiation
cell sheets
Cell Differentiation
Cell Biology
Actins
smooth muscle cells
Cell biology
Mice
Inbred C57BL
030104 developmental biology
lcsh:Biology (General)
biology.protein
Female
ACTA2
lcsh:Medicine (General)
030217 neurology & neurosurgery
Developmental Biology
Zdroj: Stem Cell Reports, Vol 13, Iss 3, Pp 499-514 (2019)
Stem Cell Reports
ISSN: 2213-6711
Popis: Summary 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.
Graphical Abstract
Highlights • ACTA2 reporter iPSC lines allow isolation of iPSC-derived SMCs • iPSC-SMCs have transcriptomic profile reminiscent of immature or synthetic SMCs • Generated iPSC-SMC cell sheets with mechanical properties in physiological ranges
In this article, Wong, Kotton, and colleagues developed murine and human iPSC lines with fluorochrome reporters for ACTA2, allowing for isolation of putative iPSC-derived smooth muscle-like populations (iPSC-SMC) with transcriptomic profiles reminiscent of immature or synthetic SMCs. The iPSC-SMCs were used to generate cell sheets with mechanical properties within physiological ranges to serve as building blocks for multi-layered tissue constructs.
Databáze: OpenAIRE
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