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Cortical Spheroid Model for Studying the Effects of Ischemic Brain Injury

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Abstract

Purpose

Ischemic brain injury occurs when there is reduced or complete disruption of blood flow to a brain region, such as in stroke or severe traumatic brain injury. Even short interruptions can lead to devastating effects including excitotoxicity and widespread cell death. Despite many decades of research, there are still very few therapeutic options for patients suffering from brain ischemia.

Methods

We developed an in vitro brain ischemia model using our previously established 3D spheroids derived from primary postnatal rat cortex. These spheroids provide an in vivo-relevant model containing a similar cellular composition to the native cortex and a cell-synthesized extracellular matrix. This model is cost-effective, highly reproducible, and can be produced in a high-throughput manner, making it an ideal candidate for screening potential therapeutics. To study the cellular and molecular mechanisms of stroke in this model, spheroids were deprived of glucose, oxygen, or both oxygen and glucose for 24 h.

Results

Both oxygen and oxygen-glucose deprived spheroids demonstrated many of the hallmarks of ischemic brain injury, including a decrease in metabolism, an increase in neural dysfunction, breakdown in the neurovascular unit, and an increase in reactive astrocytes. Pretreatment of spheroids with the antioxidant agent N-acetylcysteine (NAC) mitigated the decrease in ATP after oxygen–glucose deprivation, was partially neuroprotective, and enhanced the expression of laminin.

Conclusion

This 3D cortical spheroid model provides a platform for studying ischemic injury and has the potential for screening therapeutics.

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Data availability

Data will be made available on reasonable request.

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Acknowledgements

The authors thank Revanna Navarro, Francesca Vecchio, Austin Roy, and Dominick Calvao for assistance with data analysis and Jess Sevetson and Deanna Stueber for technical assistance and helpful discussion. The authors also thank Jason Ritt from the Carney Institute for Brain Science for assistance with statistical analysis and Geoff Williams from the Leduc Bioimaging Facility for confocal microscopy training and technical assistance.

Funding

This research was funded by a National Institute of Mental Health-funded predoctoral fellowship to RMM (T32 MH020068), a National Heart, Lung, and Blood Institute-funded predoctoral fellowship to RMM (T32 HL134625), a Brown University Sidney E. Frank Fellowship to RMM, a Bioengineering Research Partnership award National Institute of Environmental Health Sciences U01ES028184 to DHK, and the U. S. Office of Naval Research under PANTHER award number N000142112044 to DHK through Dr. Timothy Bentley.

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Authors and Affiliations

Authors

Contributions

RMM and DHK designed the research. RMM, IT, and LLL conducted research. LK and SGZ performed unpublished preliminary experiments. RMM, AL, CH, and HK analyzed data. RMM and DHK wrote the paper.

Corresponding author

Correspondence to Diane Hoffman-Kim.

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Ethics and approval

All animal procedures were conducted in accordance with the guidelines established by the NIH and approved by Brown University Institutional Animal Care and Use Committee (Protocol Number 20–07-0003).

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The authors have no competing interests to declare that are relevant to the content of this article.

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All authors have reviewed the submitted material and consent to the publication of this work.

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McLaughlin, R.M., Top, I., Laguna, A. et al. Cortical Spheroid Model for Studying the Effects of Ischemic Brain Injury. In vitro models 2, 25–41 (2023). https://doi.org/10.1007/s44164-023-00046-z

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