STAR Protocols (Dec 2022)
Generation of human elongating multi-lineage organized cardiac gastruloids
Abstract
Summary: Human elongating multi-lineage organized (EMLOC) gastruloid technology captures key aspects of trunk neurodevelopment including neural integration with cardiogenesis. We generate multi-chambered, contractile EMLOC gastruloids with integrated central and peripheral neurons using defined culture conditions and signaling factors. hiPSC colonies are primed by activating FGF and Wnt signaling pathways for co-induced lineages. EMLOC gastruloids are then initialized with primed cells in suspension culture using timed exposure to FGF2, HGF, IGF1, and Y-27632. Cardiogenesis is stimulated by FGF2, VEGF, and ascorbic acid.For complete details on the use and execution of this protocol, please refer to Olmsted and Paluh (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.