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Stem Cell Res. 2018 Aug;31:62-70. doi: 10.1016/j.scr.2018.07.008. Epub 2018 Jul 17.

Differentiation of human iPSCs into VSMCs and generation of VSMC-derived calcifying vascular cells.

Author information

1
Institute for Cardiogenetics, University of Luebeck, Germany; DZHK (German Centre for Cardiovascular Research), Partner Site Hamburg/Kiel/Luebeck, Germany; University Heart Centre Luebeck, 23562 Luebeck, Germany.
2
Institute for Cardiogenetics, University of Luebeck, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), 22525 Hamburg, Germany; DZHK (German Centre for Cardiovascular Research), Partner Site Hamburg/Kiel/Luebeck, Germany; University Heart Centre Luebeck, 23562 Luebeck, Germany.
3
Institute for Neurogenetics, University of Luebeck, 23562 Luebeck, Germany.
4
Institute for Cardiogenetics, University of Luebeck, Germany; Center for Public Health Genomics, Department of Biomedical Engineering, University of Virginia, Charlottesville, VA 22908, USA; DZHK (German Centre for Cardiovascular Research), Partner Site Hamburg/Kiel/Luebeck, Germany; University Heart Centre Luebeck, 23562 Luebeck, Germany.
5
Institute for Cardiogenetics, University of Luebeck, Germany; DZHK (German Centre for Cardiovascular Research), Partner Site Hamburg/Kiel/Luebeck, Germany; University Heart Centre Luebeck, 23562 Luebeck, Germany. Electronic address: zouhair.aherrahrou@uni-luebeck.de.

Abstract

Vascular calcification displays a major cause of death worldwide, which involve mainly vascular smooth muscle cells (VSMCs). Since 2007, there are increasing numbers of protocols to obtain different cell types from human induced-pluripotent stem cells (iPSCs), however a protocol for calcification is missing. Few protocols exist today for the differentiation of iPSCs towards VSMCs and none are known for their calcification. Here we present a protocol for the calcification of iPSC-derived VSMCs. We successfully differentiated iPSCs into VSMCs based on a modified protocol. Calcification in VSMCs is induced by a commercial StemXVivo™ osteogenic medium. Calcification was verified using Calcein and Alizarin Red S staining or Calcium assays, and molecular analyses showed enhanced expression of calcification-associated genes. The presented method could help to study genetic risk variants, using the CRISPR/Cas technology through the introduction of Knockouts or Knockins of risk variants. Finally, this method can be applied for drug screening.

PMID:
30029055
DOI:
10.1016/j.scr.2018.07.008
[Indexed for MEDLINE]
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