Generation of human elongating multi-lineage organized cardiac gastruloids

Zachary T Olmsted; Maria Belen Paredes-Espinosa; Janet L Paluh

doi:10.1016/j.xpro.2022.101898

Generation of human elongating multi-lineage organized cardiac gastruloids

STAR Protoc. 2022 Dec 16;3(4):101898. doi: 10.1016/j.xpro.2022.101898. Epub 2022 Dec 5.

Authors

Zachary T Olmsted¹, Maria Belen Paredes-Espinosa², Janet L Paluh³

Affiliations

¹ State University of New York Polytechnic Institute, College of Nanoscale Science and Engineering, Nanobioscience, Albany, NY 12203, USA; ZTO at State University of New York Downstate Medical Center, New York, NY, USA.
² State University of New York Polytechnic Institute, College of Nanoscale Science and Engineering, Nanobioscience, Albany, NY 12203, USA.
³ State University of New York Polytechnic Institute, College of Nanoscale Science and Engineering, Nanobioscience, Albany, NY 12203, USA. Electronic address: paluhj@sunypoly.edu.

Abstract

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.

Keywords: Bioinformatics; Developmental biology; Neuroscience; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ascorbic Acid
Fibroblast Growth Factor 2*
Heart
Humans
Induced Pluripotent Stem Cells*
Wnt Signaling Pathway

Substances

Fibroblast Growth Factor 2
Ascorbic Acid