A protocol to generate mature forebrain organoids from mouse embryonic stem cells is optimized and time-resolved proteomics is used to define the emergence of synaptic proteins during maturation to demonstrate that complex mature synaptic molecular architecture is present within an in vitro organoid environment.
Abstract
Neural organoids represent a promising approach for modelling neural circuits and neurological disease, yet their utility depends on whether they can recapitulate the molecular complexity of mature brain synapses. Here, we optimized a protocol to generate mature forebrain organoids from mouse embryonic stem cells and used time-resolved proteomics to define the emergence of synaptic proteins during maturation. Using fluorescence-activated synaptosome sorting coupled with mass spectrometry, we quantified the synapse-enriched proteome and found a high correlation with matched in vivo synaptic proteomes. These findings demonstrate that complex mature synaptic molecular architecture is present within an in vitro organoid environment.
The graphic presented represents a summary of the overall workflow used for creating, characterizing, and medical uses of the brain organoids derived from stem cells, and represents a detailed process of brain organoid research and its application.
H. Marei, Anwarul Hasan· Cellular and molecular neuro...· 0 citations
Despite persistent challenges related to neuronal maturation, vascularization, reproducibility, and functional integration, this review summarizes recent progress in enhancing cellular diversity, structural organization, and functional complexity within brain organoids.
Chun-Hui Duan, Yu-Kai Wang, Tie-Shan Tang et al.· Brain : a journal of neurolo...· 0 citations
Understanding human synapse structure and function at nanoscale resolution requires experimental systems that support both high-quality imaging and robust synaptogenesis. While brain tissue and primary neuronal cultures have been widely used, human induced pluripotent stem cells (iPSCs) offer key advantages: they ena...
Iman Rostami, Jana Leuenberger, Grischa Ott et al.· Cell Death & Disease· 0 citations
Organoids are stem cell-derived three-dimensional (3D) multicellular constructs that recapitulate key cytoarchitectural, molecular, and functional properties of their tissue of origin. Two fundamentally distinct strategies govern their generation: unguided approaches, which exploit intrinsic self-organization to prod...
It is shown that human cerebral organoids cultured for over six months recapitulate postnatal interneuron migration, enabling its mechanistic and genetic dissection and mechanistic analysis of previously inaccessible aspects of postnatal human brain development and their contributions to neurodevelopmental disease.
Sakurako Nagumo Wong, M. Noble, M. Fischer et al.· bioRxiv· 0 citations
