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

Publication date: August 2018Source: Stem Cell Research, Volume 31Author(s): Anja Trillhaase, Undine Haferkamp, Alexandra Rangnau, Marlon Märtens, Beatrice Schmidt, Michaela Trilck, Philip Seibler, Redouane Aherrahrou, Jeanette Erdmann, Zouhair AherrahrouAbstractVascular 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.
Source: Stem Cell Research - Category: Stem Cells Source Type: research