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Engineering 3D Cortical Spheroids for an In Vitro Ischemic Stroke Model
Eunmin Ko1, Mong Lung Steve Poon1, Eunyoung Park1
1Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, 291, Daehak-ro, Daejeon 34141, Republic of Korea.
ACS Biomaterials Science & Engineering
|July 19, 2021
Summary
Researchers developed a novel 3D brain spheroid model using ultralow attachment surfaces to mimic cerebral ischemia. This 3D model effectively recapitulates stroke responses, offering a better tool for studying brain diseases.
Area of Science:
- Neuroscience
- Biomedical Engineering
- Cell Biology
Background:
- Three-dimensional (3D) brain cell spheroids show promise for modeling brain pathophysiology.
- A 3D spheroidal model for cerebral ischemia, a condition mimicking stroke, has been lacking.
- Existing 2D cell cultures do not fully replicate the complex brain environment.
Purpose of the Study:
- To develop and investigate a 3D cortical spheroid model using ultralow attachment (ULA) surfaces for recapitulating cerebral ischemia.
- To compare the performance of ULA-surface-formed spheroids with those formed on agarose surfaces.
- To validate the 3D model's ability to mimic stroke pathophysiology through oxygen-glucose deprivation-reoxygenation (OGD-R) treatment.
Main Methods:
- Primary rat cortical cells were cultured on ultralow attachment (ULA) surfaces to form 3D spheroids.
- Comparison between 2D and 3D culture models was performed.
- Oxygen-glucose deprivation-reoxygenation (OGD-R) treatment was applied to induce ischemic conditions in the 3D spheroids.
- Gene expression analysis of stroke markers (S100B, IL-1β, MBP) and cell adhesion molecules was conducted.
Main Results:
- 3D cortical spheroids on ULA surfaces demonstrated superior performance as normal brain models compared to 2D cultures.
- ULA spheroids exhibited active cell-substrate interactions and a layered structure of neurons and astrocytes.
- OGD-R treatment in ULA spheroids successfully mimicked ischemic responses, showing upregulated stroke marker expressions.
- Agarose spheroids showed passive aggregation and different cellular arrangements compared to ULA spheroids.
Conclusions:
- ULA surface-mediated 3D cortical spheroids provide a robust model for studying cerebral ischemia and stroke.
- Cell-substrate binding activity is a critical factor in generating structurally and functionally distinct brain spheroids.
- This novel 3D model offers a valuable platform for advancing brain disease research.
