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

Anja Trillhaase; Undine Haferkamp; Alexandra Rangnau; Marlon Märtens; Beatrice Schmidt; Michaela Trilck; Philip Seibler; Redouane Aherrahrou; Jeanette Erdmann; Zouhair Aherrahrou

doi:10.1016/j.scr.2018.07.008

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

Anja Trillhaase, Undine Haferkamp, Alexandra Rangnau, Marlon Märtens, Beatrice Schmidt, Michaela Trilck, Philip Seibler, Redouane Aherrahrou, Jeanette Erdmann, Zouhair Aherrahrou^*

^*Korrespondierende/r Autor/-in für diese Arbeit

3 Zitate (Scopus)

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.

Originalsprache	Englisch
Zeitschrift	Stem Cell Research
Jahrgang	31
Seiten (von - bis)	62-70
Seitenumfang	9
ISSN	1873-5061
DOIs	https://doi.org/10.1016/j.scr.2018.07.008
Publikationsstatus	Veröffentlicht - 08.2018

Fördermittel

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 ).

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Querschnittsbereich: Medizinische Genetik

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10.1016/j.scr.2018.07.008

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title = "Differentiation of human iPSCs into VSMCs and generation of VSMC-derived calcifying vascular cells",

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{\texttrademark} 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.",

author = "Anja Trillhaase and Undine Haferkamp and Alexandra Rangnau and Marlon M{\"a}rtens and Beatrice Schmidt and Michaela Trilck and Philip Seibler and Redouane Aherrahrou and Jeanette Erdmann and Zouhair Aherrahrou",

year = "2018",

month = aug,

doi = "10.1016/j.scr.2018.07.008",

language = "English",

volume = "31",

pages = "62--70",

journal = "Stem Cell Research",

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T1 - Differentiation of human iPSCs into VSMCs and generation of VSMC-derived calcifying vascular cells

AU - Trillhaase, Anja

AU - Haferkamp, Undine

AU - Rangnau, Alexandra

AU - Märtens, Marlon

AU - Schmidt, Beatrice

AU - Trilck, Michaela

AU - Seibler, Philip

AU - Aherrahrou, Redouane

AU - Erdmann, Jeanette

AU - Aherrahrou, Zouhair

PY - 2018/8

Y1 - 2018/8

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/85049921215

U2 - 10.1016/j.scr.2018.07.008

DO - 10.1016/j.scr.2018.07.008

M3 - Journal articles

C2 - 30029055

AN - SCOPUS:85049921215

SN - 1873-5061

VL - 31

SP - 62

EP - 70

JO - Stem Cell Research

JF - Stem Cell Research

ER -

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

Abstract

Fördermittel

UN SDGs

Strategische Forschungsbereiche und Zentren

Dokumente

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