Development and Characterization of Human Cerebral Organoids: An Optimized Protocol

Cell Transplantation
Abraam M Yakoub, Mark Sadek

Abstract

Studies of human neurodevelopmental disorders and stem cell-based regenerative transplants have been hampered by the lack of a model of the developing human brain. Stem cell-derived neurons suffer major limitations, including the ability to recapitulate the 3-dimensional architecture of a brain tissue and the representation of multiple layers and cell types that contribute to the overall brain functions in vivo. Recently, cerebral organoid technology was introduced; however, such technology is still in its infancy, and its low reproducibility and limitations significantly reduce the reliability of such a model as it currently exists, especially considering the complexity of cerebral-organoid protocols. Here we have tested and compared multiple protocols and conditions for growth of organoids, and we describe an optimized methodology, and define the necessary and sufficient factors that support the development of optimal organoids. Our optimization criteria included organoids' overall growth and size, stratification and representation of the various cell types, inter-batch variability, analysis of neuronal maturation, and even the cost of the procedure. Importantly, this protocol encompasses a plethora of technical tips that all...Continue Reading

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Citations

Apr 12, 2019·Journal of Cellular Physiology·Alan W Leung, James Y H Li
Jun 5, 2019·Cell Transplantation·Abraam M Yakoub, Mark Sadek
Feb 11, 2020·Frontiers in Neuroscience·Marco TjakraGuixue Wang
Mar 21, 2019·Frontiers in Neuroscience·Daniele PoliArti Ahluwalia
Apr 27, 2019·Frontiers in Cellular Neuroscience·Dario PacittiBridget E Bax
Nov 4, 2020·Scientific Reports·Divya SriramVegesna Radha

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Methods Mentioned

BETA
Reverse-transcription Polymerase Chain Reaction
PCR
confocal microscopy

Software Mentioned

RQ Manager

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