In Vitro Stroke Models & Assay Services

IN VITRO STROKE MODELS

Stroke remains one of the leading causes of death and long-term disability worldwide, driven by complex pathophysiological cascades including ischemic energy failure, excitotoxicity, oxidative stress, neuroinflammation, and programmed cell death. Robust in vitro models that faithfully recapitulate key stroke-related pathologies are essential for accelerating mechanistic understanding and therapeutic discovery.

Our platform employs a panel of well-characterized cellular models that target distinct yet interconnected mechanisms of ischemic brain injury. From oxygen-glucose deprivation (OGD) simulating ischemia/reperfusion, to glutamate-induced excitotoxicity modeling neuronal overstimulation, and LPS-activated microglial inflammation reflecting secondary immune responses — each model is paired with multi-parameter phenotypic and molecular readouts to deliver mechanistic insights and pharmacological validation.Our assay portfolio includes CCK-8 cell viability assay, LDH cytotoxicity assay, cellular ROS detection, Griess assay for nitric oxide quantification, immunofluorescence staining, ELISA for cytokine profiling, and neuronal morphological analysis — enabling comprehensive evaluation of compound efficacy across cell survival, oxidative stress, inflammatory response, apoptosis, and neurite integrity.

These models serve as powerful, scalable tools for compound screening, mechanism-of-action (MoA) studies, and neuroprotective candidate validation prior to in vivo translation.

MODEL SELECTION

Mechanism-driven in vitro model selection for stroke drug discovery

Modeling Method	Primary pathological mechanism modeled	Commonly used cell type	Key molecular pathways	Main phenotypic readout	Optimal use case
OGD-induced	Ischemic substrate deprivation; energy failure, oxidative stress, mitochondrial dysfunction, ER stress, inflammasome activation, apoptosis or parthanatos	SH-SY5Y neuroblastoma cells, primary hippocampus neuron	ATP depletion, ROS, mitochondrial depolarization, ER stress ATF4/ATF6/PERK/IRE1, PARP-1/PAR/AIF, NF-κB, NLRP3, Nrf2/HO-1, AMPK	MTT/CCK-8, LDH, trypan blue, ROS, MDA/GSH, JC-1 mitochondrial potential	Screening compounds against ischemia-like neuronal energy failure; testing mitochondrial, ER-stress, parthanatos, inflammasome, and antioxidant mechanisms
Glutamate-induced	Excitotoxic and oxidative neuronal injury without necessarily reproducing full ischemic deprivation	SH-SY5Y cells; primary cortical or hippocampal neurons	ROS, lipid and protein oxidation, mitochondrial dysfunction, Bax/Bcl-2, cytochrome c, caspase-3, JNK and p38 MAPK; receptor dependence must be validated in the chosen clone and differentiation state	Viability, LDH, ROS, MDA, protein carbonyls, SOD/catalase, mitochondrial membrane potential, Annexin V/PI, Bax/Bcl-2/cleaved caspase-3	Isolating excitotoxic/oxidative injury and testing mitochondrial or MAPK-targeted neuroprotection; not sufficient alone to infer protection from full ischemia
LPS-induced	Microglial inflammatory amplification and innate immune activation; models secondary neuroinflammation rather than ischemic energy failure	Murine BV2 microglia	TLR4/MyD88/IRAK, IκBα/NF-κB, ERK/JNK/p38 MAPK, iNOS/NO, COX-2/PGE2, IL-1β, TNF-α, IL-6, miR-155, STAT3 and NLRP3 depending on protocol	Griess NO, PGE2, TNF-α, IL-1β, IL-6, microglial morphology, neuronal death in coculture	Testing anti-inflammatory mechanisms, microglial signaling, and immune-mediated neuronal injury; should be paired with OGD or in vivo models for stroke relevance

VALIDATED MODEL LIBRARY

Explore the model-specific validation workflow

Select a model below to review its biological rationale, experimental design, efficacy readouts, and representative data.

01OGD/R-Induced SH-SY5Y Cell Model

In the oxygen-glucose deprivation/reoxygenation (OGD/R) mode, we offer CCK-8 cell viability assay and cellular ROS detection to evaluate neuroprotective effects of test compounds. The OGD/R model successfully reproduced ischemia/reperfusion-induced neuronal injury, characterized by reduced cell viability and excessive oxidative stress. The Edaravone/Dexborneol treatment effectively improved cell survival and attenuated oxidative stress.

CCK-8 cell viability and cellular ROS detection in OGD/R model

Mechanistically, OGD/R activated apoptosis-associated signaling by Caspase-3 immunofluorescence and inflammatory responses by IL-1β ELISA, whereas Edaravone/Dexborneol effectively suppressed both neuronal apoptosis and the inflammatory cascade — confirming the model's utility for evaluating multi-target neuroprotective agents.

Caspase-3 immunofluorescence and IL-1β detection in OGD/R model

02LPS-Induced BV2 Cell Model

This model utilizes lipopolysaccharide (LPS) to activate murine BV2 microglia, simulating the secondary neuroinflammatory response that exacerbates ischemic brain damage. Concentration gradient screening was carried out via the CCK-8 cell viability assay, and 1 μg/mL LPS was selected as the optimal concentration for establishing the inflammatory model.

LPS concentration gradient screening in BV2 cells

This model exhibited robust inflammatory responses, with increased ROS levels measured by fluorescence imaging, elevated NO production quantified via the Griess assay, and upregulated IL-6 and TNF-α secretion detected by ELISA. Dexamethasone significantly attenuated these pathological changes, with dose-dependent inhibition of IL-6 and TNF-α, validating the model as a reliable platform for anti-inflammatory drug screening.

Inflammatory responses in LPS-induced BV2 model

03Glutamate-Induced SH-SY5Y Cell Model

Glutamate excitotoxicity is a major driver of neuronal death during cerebral ischemia. This model uses glutamate to induce excitotoxic injury in SH-SY5Y neuroblastoma cells. Through concentration gradient screening via the CCK-8 cell viability assay, 20 mM glutamate was identified as the optimal modeling concentration. This condition stably induces neuronal injury and provides a robust platform for evaluating neuroprotective compounds against glutamate-induced excitotoxicity.

Glutamate concentration gradient screening in SH-SY5Y cells

The glutamate-induced SH-SY5Y cell model exhibited robust excitotoxic responses, including increased oxidative stress by ROS detection and elevated LDH release by LDH cytotoxicity assay. Treatment with 7-BIO, Resveratrol, and Donepezil significantly attenuated glutamate-induced cytotoxicity, demonstrating the model's reliability and strong pharmacological responsiveness for excitotoxicity-targeted neuroprotection studies.

Excitotoxic responses in glutamate-induced SH-SY5Y model

04Glutamate-Induced Primary Hippocampal Neuron Injury Model

Building on the SH-SY5Y excitotoxicity model, this study extends to primary hippocampal neurons to capture more physiologically relevant morphological and structural responses to glutamate-induced injury.The glutamate-induced primary hippocampal neuron model was successfully established, characterized by significant decreases in total neurite length, maximum neurite length, neurite count, and branch points by neuronal morphological analysis. These findings reflect impaired neuronal morphology and complexity, providing a sensitive readout for structural neuroprotection and neuronal network integrity in the context of excitotoxic stroke injury.

Neuronal morphology in glutamate-induced primary hippocampal neuron model

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