Integrating central and peripheral neurons in elongating multi-lineage-organized gastruloids

Abstract

Human stem cell technologies including self-assembling 3D tissue models provide unprecedented access to early neurodevelopment and enable fundamental insights into neuropathologies. Gastruloid models have yet to be used to investigate the developing nervous system. Here we generate elongating multi-lineage-organized (EMLO) gastruloids with trunk identity that co-develop central and peripheral nervous system (CNS, PNS) correlates. We track migrating neural crest cells that differentiate to form peripheral neurons integrated with an upstream spinal cord region. This follows initial EMLO polarization events, and is coordinated with primitive gut tube elongation and cardiomyocyte differentiation. By immunofluorescence of multi-lineage and functional biomarkers, we evaluate EMLOs over a twenty-two day period, and apply them to investigate the impact of mu opioid receptor modulation on neuronal activity. This comprehensive study demonstrates a novel combined CNS-PNS model of early organogenesis and integration events in the trunk to benefit human biomedical research.