Murine cerebral organoids develop network of functional neurons and hippocampal brain region identity

Abstract

Summary Brain organoids are in vitro three-dimensional (3D) self-organized neural structures, which can enable disease modeling and drug screening. However, their use for standardized large-scale drug screening studies is limited by their high batch-to-batch variability, long differentiation time (10–20 weeks), and high production costs. This is particularly relevant when brain organoids are obtained from human induced pluripotent stem cells (iPSCs). Here, we developed, for the first time, a highly standardized, reproducible, and fast (5 weeks) murine brain organoid model starting from embryonic neural stem cells. We obtained brain organoids, which progressively differentiated and self-organized into 3D networks of functional neurons with dorsal forebrain phenotype. Furthermore, by adding the morphogen WNT3a, we generated brain organoids with specific hippocampal region identity. Overall, our results showed the establishment of a fast, robust and reproducible murine 3D in vitro brain model that may represent a useful tool for high-throughput drug screening and disease modeling.

Revolutionizing Disease Modeling: The Emergence of Organoids in Cellular Systems

“…Organoids are a technology with many potential benefits and applications, but there are also several limitations that must be considered. The generation of organoids has a high production cost, mainly when obtained from iPSCs, because of material price, iPSCs maintenance, long differentiation and maturation time of organoids [ 284 ], depending on protocol differentiation and specificity (vascularization and cell type differentiation) [ 143 , 285 , 286 , 287 , 288 ]. Additionally, the lack of vascularization in organoids limits the supply of oxygen and nutrients compared with in vivo cells near capillaries [ 145 , 150 , 289 ].…”

“…Furthermore, the variability of organoids in batch-to-batch production is caused by stem cell organization mechanisms, the cell types of formation control absence, the variability of position of the organ regions, and the organoid size and shape difference. Additionally, the differentiation and growth factors available in the culture system limit not only the unbiased drug screening studies but also the modeling diseases in quantitative studies and the standardization of scientific research [ 143 , 144 , 150 , 284 , 299 ]. Hence, the organoids technology is valuable for studying human disorders, although there are restrictions that must be overcome, such as low homogeneity and high costs [ 21 , 145 ].…”

Bilirubin-Induced Neurological Damage: Current and Emerging iPSC-Derived Brain Organoid Models

“…Cerebral organoid research has significantly expanded within the last decade with the introduction of more complexity and specificity [ 157 ]. Relying on small molecules, individual brain-region-specific organoids can be generated by growth-factor-based manipulation, which consequently determine the cellular identities such as cerebral organoids with choroid plexus, hippocampus, retina and striatum [ 158 , 159 , 160 , 161 ]. The generation of organoids by co-culturing different cells is another advanced and innovative approach to enhance the model complexity and to investigate the cell–cell and cell–matrix interplays in a 3D environment during human brain development and disease [ 162 , 163 , 164 ].…”

Technical advances in pluripotent stem cell-derived and tumorigenic organoids

“…After developing cerebral organoids containing various brain regions, subsequent studies focused on generating region-specific spheroids [8,9]. Organoids or region-specific spheroids have been developed for other brain regions, such as the thalamus [10,11], choroid plexus [12], midbrain [13,14], cerebellum [15,16], and hippocampus [17][18][19]. Other advances have been made to generate brain organoid models more closely resembling the in vivo organ.…”