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.
| Original language | English |
|---|---|
| Journal | Stem Cell Research |
| Volume | 31 |
| Pages (from-to) | 62-70 |
| Number of pages | 9 |
| ISSN | 1873-5061 |
| DOIs | |
| Publication status | Published - 08.2018 |
Funding
This study was funded by a grant from the Fondation Leducq ( 12CVD02 ; CADgenomics: Understanding Coronary Artery Disease Genes). P.S. is supported by the German Research Foundation ( FOR2488 ).
Research Areas and Centers
- Research Area: Medical Genetics
