Development of an in vitro Blood Clot Model to Simulate Intracerebral Hemorrhage for Neuroprotective Drug Screening
Alyssa Han
Chadwick International School, Incheon, South Korea
Publication date: November 20, 2025
Chadwick International School, Incheon, South Korea
Publication date: November 20, 2025
DOI: http://doi.org/10.34614/JIYRC2025II08
ABSTRACT
Intracerebral hemorrhage (ICH), a severe form of stroke caused by bleeding within the brain tissue, remains a growing global health concern with limited treatment options. Traditional ICH models often rely on animal studies or computational simulations, which pose ethical, financial, and experimental limitations. To address this gap, we developed a simple, reproducible in vitro model of ICH using agarose gel to simulate blood clot formation and evaluate its toxic effects on human brain cells. Agarose—a biocompatible, transparent hydrogel—was mixed with either phosphate-buffered saline (PBS) or human whole blood at various concentrations (1%, 2%, and 4%) to form clot-like structures. These were co-cultured with A172 human astrocyte cells for seven days. Fluorescence-based live/dead cell assays demonstrated that higher agarose concentrations impeded blood diffusion and significantly reduced brain cell viability, with the 4% agarose-blood condition resulting in a viability drop to 51.3% (p = 0.0001). This model provides a cost-effective and ethically sound alternative to in vivo methods, allowing precise control over clot parameters. It offers a promising platform for simulating ICH pathology and screening neuroprotective drugs, while acknowledging limitations such as the absence of hemodynamic forces and multi-cellular interactions.
Intracerebral hemorrhage (ICH), a severe form of stroke caused by bleeding within the brain tissue, remains a growing global health concern with limited treatment options. Traditional ICH models often rely on animal studies or computational simulations, which pose ethical, financial, and experimental limitations. To address this gap, we developed a simple, reproducible in vitro model of ICH using agarose gel to simulate blood clot formation and evaluate its toxic effects on human brain cells. Agarose—a biocompatible, transparent hydrogel—was mixed with either phosphate-buffered saline (PBS) or human whole blood at various concentrations (1%, 2%, and 4%) to form clot-like structures. These were co-cultured with A172 human astrocyte cells for seven days. Fluorescence-based live/dead cell assays demonstrated that higher agarose concentrations impeded blood diffusion and significantly reduced brain cell viability, with the 4% agarose-blood condition resulting in a viability drop to 51.3% (p = 0.0001). This model provides a cost-effective and ethically sound alternative to in vivo methods, allowing precise control over clot parameters. It offers a promising platform for simulating ICH pathology and screening neuroprotective drugs, while acknowledging limitations such as the absence of hemodynamic forces and multi-cellular interactions.
