Co‐Differentiation of Human Pluripotent Stem Cells‐Derived Cardiomyocytes and Endothelial Cells from Cardiac Mesoderm Provides a Three‐Dimensional Model of Cardiac Microtissue. (13th February 2018)

• Record Type: Journal Article
• Authors: Giacomelli, Elisa; Bellin, Milena; Orlova, Valeria V.; Mummery, Christine L.
• Editors: Dracopoli, Nicholas C.; Haines, Jonathan L.; Korf, Bruce R.; Morton, Cynthia C.; Seidman, Christine E.; Seidman, J.G.; Smith, Douglas R.
• Is Part Of: Current protocols in human genetics. Volume 95(2017)
• Page Start: 21.9.1
• Page End: 21.9.22
• Publication Date: 2018-02-13
• Subjects: human pluripotent stem cell‐derived cardiomyocytes -- human pluripotent stem cell‐derived endothelial cells -- cardiac mesoderm -- cardiac microtissues -- three‐dimensional culture model; Human genetics -- Laboratory manuals; Genetic Techniques; Human genetics; Laboratory manuals; 599.935028
• DOI: 10.1002/cphg.46
• Languages: English
• ISSNs: 1934-8258

Abstract

Abstract: The formation of cardiac mesodermal subtypes is highly regulated in time and space during heart development. In vitro models based on human pluripotent stem cells (hPS cells) provide opportunities to study mechanisms underlying fate choices governing lineage specification from common cardiovascular progenitors in human embryos. The generation of cardiac endothelial cells in particular allows the creation of complex models of cardiovascular disorders in which either cardiomyocytes or endothelial cells are affected. Here, a protocol for co‐differentiation of cardiomyocytes and endothelial cells from cardiac mesoderm using hPS cells is described. Precise details for the enrichment of each cell population from heterogeneous‐differentiated cultures, a description of how to maintain and dissociate enriched cardiomyocytes, and the expansion and cryopreservation of enriched endothelial cells are all provided. The generation and culture of three‐dimensional cardiac microtissues from these cell populations is described and guidelines for the characterization of microtissues by immunofluorescent staining and re‐plating for downstream applications are provided. © 2017 by John Wiley & Sons, Inc.