Generation of spatial-patterned early-developing cardiac organoids using human pluripotent stem cells

• Published in: Nature protocols Vol. 13; no. 4; pp. 723 - 737
• Main Authors: Hoang, Plansky, Wang, Jason, Conklin, Bruce R, Healy, Kevin E, Ma, Zhen
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.04.2018; Nature Publishing Group
• Subjects: 13/100; 13/107; 631/136/1660/1986; 631/136/2060; 631/1647/245; 631/443/592/2726; 631/532/1360; Analytical Chemistry; Biological products; Biological Techniques; Biomaterials; Biomedical materials; Cardiomyocytes; Computational Biology/Bioinformatics; Coronary artery disease; Developmental biology; Differentiation; Drug discovery; Embryogenesis; Embryonic growth stage; Fetuses; Health aspects; Heart; Heart - growth & development; Heart diseases; Hereditary diseases; Humans; Induced Pluripotent Stem Cells; Life Sciences; Medical research; Microarrays; Models, Biological; Morphogenesis; Organ Culture Techniques - methods; Organic Chemistry; Organogenesis; Organoids; Organoids - growth & development; Pattern formation; Photolithography; Pluripotency; protocol; Self-assembly; Stem cells
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/nprot.2018.006

This protocol describes the generation of early-developing cardiac organoids from human pluripotent stem cells. Geometric confinement of the hiPSCs drives spatial organization of the cells from a 2D layer into 3D cardiac microchambers. The creation of human induced pluripotent stem cells (hiPSCs) has provided an unprecedented opportunity to study tissue morphogenesis and organ development through 'organogenesis-in-a-dish'. Current approaches to cardiac organoid engineering rely on either direct cardiac differentiation from embryoid bodies (EBs) or generation of aligned cardiac tissues from predifferentiated cardiomyocytes from monolayer hiPSCs. To experimentally model early cardiac organogenesis in vitro , our protocol combines biomaterials-based cell patterning with stem cell organoid engineering. 3D cardiac microchambers are created from 2D hiPSC colonies; these microchambers approximate an early-development heart with distinct spatial organization and self-assembly. With proper training in photolithography microfabrication, maintenance of human pluripotent stem cells, and cardiac differentiation, a graduate student with guidance will likely be able to carry out this experimental protocol, which requires ∼3 weeks. We envisage that this in vitro model of human early heart development could serve as an embryotoxicity screening assay in drug discovery, regulation, and prescription for healthy fetal development. We anticipate that, when applied to hiPSC lines derived from patients with inherited diseases, this protocol can be used to study the disease mechanisms of cardiac malformations at an early stage of embryogenesis.