Abstract
Trunk-biased human gastruloids provide the ability to couple developmentally relevant spinal neurogenesis and organ morphogenesis via spatiotemporal self-organization events from derivatives of the three germ layers. The multi-lineage nature of gastruloids provides the full complexity of regulatory signaling cues that surpasses directed organoids and lays the foundation for an ex vivo self-evolving system. Here we detail two distinct protocols for trunk-biased gastruloids from an elongated, polarized structure with coordinated organ-specific neural patterning. Following an induction phase to caudalize iPSCs to trunk phenotype, divergent features of organogenesis and end-organ innervation yield separate models of enteric and cardiac nervous system formation. Both protocols are permissive to multi-lineage development and allow the study of neural integration events within a native, embryo-like context. We discuss the customizability of human gastruloids and the optimization of initial and extended conditions that maintain a permissive environment for multi-lineage differentiation and integration.
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Acknowledgments
This work was supported by multiple New York state awards. EMLO work was funded by the NY State Department of Health (NYS DOH) Spinal Cord Injury Research Board (NYSCIRB), Projects to Accelerate Research Translation (PART) award C33278GG and SUNY Polytechnic SEED award 917035-21 and used published lines developed through previous New York State stem cell research (NYSTEM) funding. The EMLOC research was supported by SUNY Polytechnic SEED award 917035 and CATN2 award 1180838-1-92476. The scRNAseq was performed at the SUNY Buffalo Genomics and Bioinformatics Core.
Conflicts of Interest
The authors declare the following potential conflict of interest on patent pending EMLOC technology: 63/311,498, “A combined human gastruloid model for cardiogenesis and neurogenesis.”
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Olmsted, Z.T., Paredes-Espinosa, M.B., Paluh, J.L. (2023). Embryonic Spinal Cord Innervation in Human Trunk Organogenesis Gastruloids: Cardiac Versus Enteric Customization and Beyond. In: Zernicka-Goetz, M., Turksen, K. (eds) Embryo Models In Vitro. Methods in Molecular Biology, vol 2767. Humana, New York, NY. https://doi.org/10.1007/7651_2023_491
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DOI: https://doi.org/10.1007/7651_2023_491
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