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

Stem Cell Research
Anja TrillhaaseZouhair Aherrahrou

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.

Citations

Nov 17, 2020·Frontiers in Cell and Developmental Biology·Hongorzul DavaapilSanjay Sinha
Dec 2, 2020·Medical Hypotheses·Stuart D FerrellSanford H Barsky

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