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more accurately. For example, Nzou et al. developed 3D The permeability of the compound can be quantified
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NVU organoids using a hanging-drop culture method. Such by measuring the mean fluorescence intensity through
organoids, which comprise neurons, oligodendrocytes, confocal fluorescence z-stack images. This method
astrocytes, microglia, human brain microvascular ECs, and allows for rapid testing of different compounds, such
pericytes, were exposed to hypoxic conditions to simulate as angiopep-2, phosphatidylinositol 3-kinase inhibitor
stroke-induced BBB dysfunction. This model successfully BKM120, and dabrafenib. In addition, V-Organoids can
recapitulated features such as increased permeability, be used to screen anti-inflammatory compounds and
inflammatory responses, and oxidative stress, providing identify agents that restore BBB integrity and counteract
a physiologically relevant platform for stroke research. the invasive properties of GBM cells. V-Organoids can
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Although studies on 3D stroke-related organoid models simulate the crucial aspects of BBB, providing a low-cost
are limited, advancements in organoid technology suggest approach to evaluate the brain-permeable molecules for a
a promising future for stroke modeling. wide range of diseases. Future research may benefit from the
The presence of functional vasculature in V-Organoids ongoing refinement of V-Organoids, particularly through
provides a more physiologically relevant platform for the incorporation of vascular and immune components to
studying brain cancers, particularly in modeling the blood– more accurately simulate in vivo conditions.
brain tumor barrier, which arises as the BBB undergoes a 4.3. Regeneration applications of vascularized BOs
pathological transformation during tumor progression.
Unlike traditional organoid models, which lack perfusion Current cell transplantation therapies for brain injuries face
and vascular remodeling, V-Organoids closely mimic the limitations in regenerating multiple damaged cell types.
tumor microenvironment by integrating dynamic vascular Most cell transplantation research has employed single
networks. This allows researchers to investigate tumor- NSCs or neuronal cell types to repair brain injuries, which
endothelial interactions, angiogenic signaling, and barrier may be insufficient for regenerating the wide array of cell
permeability changes in GBM. This vascular complexity types lost during injury. 182,187 BOs, however, contain a diverse
is critical for evaluating anti-angiogenic therapies, which range of neural cell types, presenting a rich source of cells
target tumor-induced neovascularization, as well as for transplantation, brain injury repair, and regeneration.
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for testing BBB-penetrating drugs that must cross the Wang et al. assessed the effect of COs’ transplantation in
blood–brain tumor barrier to reach malignant cells. By a rat model of ischemic stroke (Figure 10A). Their findings
providing a controlled yet physiologically relevant in vitro demonstrate that transplanted COs could differentiate
system, V-Organoids enable high-fidelity drug screening into neurons, glial cells, and other cortical cell types,
and therapeutic development, bridging the gap between effectively mimicking cortical regeneration. This process
preclinical research and clinical applications in brain supports neurogenesis, synaptic reconstruction, and axonal
cancer treatment. Moreover, V-Organoids have enabled regrowth, thus enabling the formation of functional neural
researchers to more accurately replicate the complex circuits within the damaged brain region. Meanwhile, Cao
interactions between GBM cells and the neurovascular et al. 13,189 showed that COs’ transplantation is more effective
environment. Grebenyuk and Ranga used V-Organoids in repairing structural damage compared to individual
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to demonstrate how GBM cells may compromise BBB neural stem cell transplants, providing a novel strategy
structure, resulting in alterations of TJ proteins and for stroke rehabilitation. In their ischemic stroke mouse
increased permeability, thus mimicking the in vivo tumor model, transplanted medial ganglionic eminence organoids
microenvironment. Moreover, the integration of microglia survived in the infarcted cortex, differentiated into specific
into V-Organoids has allowed for the investigation of cross- cortical neuron subtypes, and projected axons to establish
talk between GBM cells and immune cells, offering insights connections with host neural circuits, leading to significant
into how tumors modulate the immune response to improvements in sensorimotor functions. Furthermore,
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support their growth. This multifaceted approach advances Cao et al. transplanted COs enriched in GABAergic
our understanding of GBM biology, including tumor interneurons into stroke-affected mouse cortices. These
angiogenesis, immune evasion, and BBB disruption, all of organoids integrated effectively within the damaged cortex,
which are critical for developing more effective therapies differentiated robustly, and restored sensorimotor function
for brain cancers. 185 in the stroke-affected mice. These observations suggest that
In addition to disease modeling, V-Organoids have a specific types of organoids, particularly those containing
wide range of applications in CNS drug development. As inhibitory neurons, can facilitate recovery post-stroke by
previously mentioned, the treatment of CNS diseases may replenishing lost neuronal populations and reconstructing
functional neural circuits (Figure 10B).
be hampered by the BBB. To address this, Bergmann
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et al. discussed how BBB organoids can be utilized Given most brain injuries result from ischemia,
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to determine whether compounds can cross the BBB. vascularization in BOs could significantly enhance their

Volume 1 Issue 2 (2025) 23 doi: 10.36922/or.8162