Stem Cell Res. 2018 Jul 17;31:62-70. doi: 10.1016/j.scr.2018.07.008. [Epub ahead of print]

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

Trillhaase A¹, Haferkamp U², Rangnau A¹, Märtens M¹, Schmidt B¹, Trilck M³, Seibler P³, Aherrahrou R⁴, Erdmann J¹, Aherrahrou Z⁵.

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.