Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids.
- Authors
- Salick, Max R; Wells, Michael F; Eggan, Kevin; Kaykas, Ajamete
- Year
- 2017
- Journal
- Journal of visualized experiments : JoVE
- PMID
- 28994790
- DOI
- 10.3791/56404
- PMCID
- PMC5752258
The recent emergence of Zika virus (ZIKV) in susceptible populations has led to an abrupt increase in microcephaly and other neurodevelopmental conditions in newborn infants. While mosquitos are the main route of viral transmission, it has also been shown to spread via sexual contact and vertical mother-to-fetus transmission. In this latter case of transmission, due to the unique viral tropism of ZIKV, the virus is believed to predominantly target the neural progenitor cells (NPCs) of the developing brain. Here a method for modeling ZIKV infection, and the resulting microcephaly, that occur when human cerebral organoids are exposed to live ZIKV is described. The organoids display high levels of virus within their neural progenitor population, and exhibit severe cell death and microcephaly over time. This three-dimensional cerebral organoid model allows researchers to conduct species-matched experiments to observe and potentially intervene with ZIKV infection of the developing human brain. The model provides improved relevance over standard two-dimensional methods, and contains human-specific cellular architecture and protein expression that are not possible in animal models.
LLM interpretation
This figure consists of five bright-field microscopy images (A-E) showing cell morphology and distribution. Panels A, B, and C provide low-magnification views with a 1mm scale bar, while panels D and E provide higher-magnification views with a 200µm scale bar. Panel E includes white arrowheads pointing to specific cellular features or cells migrating between larger cell clusters.
LLM interpretation
Figure A is a schematic flow diagram illustrating a multi-step protocol for the detachment, counting, and seeding of pluripotent stem cells (PSCs) into a ULA U-bottom 96-well plate. Figures B through G are microscopy images showing the morphological development of a cellular aggregate over a time course from day 8 (D8) to day 30 (D30). The aggregates increase in size and complexity over time, with black arrowheads in F and G highlighting specific structural features emerging by D24 and D30. A 1mm scale bar is provided at the bottom.
LLM interpretation
This figure consists of a schematic diagram and a series of microscopy images. Panel A illustrates a procedural workflow involving a pipette and a blue spherical object in a vial. Panels B through E show the morphological evolution of a spherical sample over time, comparing "Day 6" (B, C) to "Day 18" (D, E), with a 1mm scale bar provided in panel B.
LLM interpretation
This figure presents immunofluorescence microscopy images of brain organoids at two time points: Day 25 (panels A-E') and Day 108 (panels F-I). At Day 25, staining shows the distribution of DAPI (blue), phVim (green), TBR2 (red), and MAP2 (white), with a merged high-magnification inset (E') labeling the ventricular zone (VZ), subventricular zone (SVZ), intermediate zone (IZ), and cortical plate (CP). At Day 108, the organoid is stained for DAPI (blue), GFAP (red), and MAP2 (white), with a final merged image (I) showing the spatial overlap of these markers.
LLM interpretation
This figure consists of microscopy images and a corresponding bar chart analyzing the effect of ZIKV-PR infection on organoid size. Panel A shows a time-course (Day 0, 3, and 6) of organoids under three conditions: Mock, MOI 0.1, and MOI 10. Panel B is a bar chart showing a significant decrease in the two-dimensional area ($\text{mm}^2$) of organoids at 6 dpi as the multiplicity of infection (MOI) increases, with statistical significance indicated by asterisks (* and **). Panels C and D provide higher-magnification images of organoid edges with $100\text{ }\mu\text{m}$ scale bars.
LLM interpretation
This figure consists of four fluorescence microscopy images (A-D) of a tissue section, with a 200 $\mu$m scale bar in panel C. Panel A shows DAPI staining (blue) for nuclei, panel B shows 4G2 staining (green) with specific clusters indicated by white arrowheads, and panel C shows MAP2 staining (red). Panel D is a merged image showing the colocalization of the three markers.
LLM interpretation
This figure consists of three rows (A, B, C) of multi-channel immunofluorescence microscopy images. Each row shows four panels: DAPI (blue) for nuclei, 4G2 (green), CC3 (red), and a merged image of all three channels. There is a visible increase in the intensity and distribution of both 4G2 and CC3 signals from row A to row C, with a scale bar of 100µm provided in the bottom-left panel.
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| Applications and research trends in organoid based infectious disease models. | Ryu JO et al. | — | 2025 | → |
| Congenital Zika virus infection in laboratory animals: a comparative review highlights translational studies on the maternal-foetal interface. | Gardinali NR et al. | — | 2025 | → |
| The Application and Challenges of Brain Organoids in Exploring the Mechanism of Arbovirus Infection. | Cui B et al. | — | 2025 | → |
| Biallelic loss-of-function variants in CACHD1 cause a novel neurodevelopmental syndrome with facial dysmorphism and multisystem congenital abnormalities. | Scala M et al. | — | 2024 | → |
| <i>REST</i> and <i>RCOR</i> genes display distinct expression profiles in neurons and astrocytes using 2D and 3D human pluripotent stem cell models. | Maksour S et al. | — | 2024 | → |
| <i>REST</i>and<i>RCOR</i>genes display distinct expression profiles in neurons and astrocytes using 2D and 3D human pluripotent stem cell models | Maksour S et al. | — | 2024 | — |
| Visualization of incrementally learned projection trajectories for longitudinal data. | Malepathirana T et al. | — | 2024 | → |
| 3D engineered tissue models for studying human-specific infectious viral diseases. | Hwang KS et al. | — | 2023 | → |
| iPSC-derived three-dimensional brain organoid models and neurotropic viral infections. | Swingler M et al. | — | 2023 | → |
| Natural variation in gene expression and viral susceptibility revealed by neural progenitor cell villages. | Wells MF et al. | — | 2023 | → |
| Alternative Methods to Current In Vivo Procedures to Address the 3Rs Tenet in Rabies Proficiency Testing. | Zorzan M et al. | — | 2022 | → |
| Human Brain Organoids as Models for Central Nervous System Viral Infection. | Depla JA et al. | — | 2022 | → |
| Human mini brains and spinal cords in a dish: Modeling strategies, current challenges, and prospective advances. | Kofman S et al. | — | 2022 | → |
| Making neurons, made easy: The use of Neurogenin-2 in neuronal differentiation. | Hulme AJ et al. | — | 2022 | → |
| Modeling infectious diseases of the central nervous system with human brain organoids. | Priyathilaka TT et al. | — | 2022 | → |
| Using 2D and 3D pluripotent stem cell models to study neurotropic viruses. | LaNoce E et al. | — | 2022 | → |
| Advancing Drug Discovery for Neurological Disorders Using iPSC-Derived Neural Organoids. | Costamagna G et al. | — | 2021 | → |
| Expression of Endogenous Angiotensin-Converting Enzyme 2 in Human Induced Pluripotent Stem Cell-Derived Retinal Organoids. | Ahmad Mulyadi Lai HI et al. | — | 2021 | → |
| Using our mini-brains: cerebral organoids as an improved cellular model for human prion disease. | Groveman BR et al. | — | 2020 | → |
| All Together Now: Modeling the Interaction of Neural With Non-neural Systems Using Organoid Models. | Chukwurah E et al. | — | 2019 | → |
| Sporadic Creutzfeldt-Jakob disease prion infection of human cerebral organoids. | Groveman BR et al. | — | 2019 | → |
| Using human stem cells as a model system to understand the neural mechanisms of alcohol use disorders: Current status and outlook. | Scarnati MS et al. | — | 2019 | → |
| Microglia Increase Inflammatory Responses in iPSC-Derived Human BrainSpheres. | Abreu CM et al. | — | 2018 | → |