Generation of human elongating multi-lineage organized cardiac gastruloids

• Published in: STAR protocols 2022-12, Vol.3 (4), p.101898-101898, Article 101898
• Main Authors: Olmsted, Zachary T., Paredes-Espinosa, Maria Belen, Paluh, Janet L.
• Format: Article
• Language: English
• Subjects: Ascorbic Acid; Bioinformatics; Developmental biology; Fibroblast Growth Factor 2; Heart; Humans; Induced Pluripotent Stem Cells; Neuroscience; Organoids; Protocol; Stem Cells; Wnt Signaling Pathway
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101898

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 [Display omitted] •2D hiPSCs primed for trunk mesoderm, endoderm, and neural differentiation•2D to 3D aggregate formation with committed neural and cardiac polarized domains•Embryo-like cardiogenesis, heart tube-to-chamber morphogenesis, and specialization•Model of trunk central and peripheral neurogenesis integrated with cardiogenesis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.