Hindbrain Organoid Modeling Service

Introduction

The hindbrain, a foundational region of the brainstem, plays a pivotal role in governing essential life processes such as breathing, coordinating movement, and regulating the autonomic nervous system. This intricate area encompasses structures like the pons, medulla oblongata, and cerebellum. Notably, it is home to the raphe nuclei – clusters of neurons that produce serotonin (5-HT), a crucial neurotransmitter modulating mood, sleep patterns, appetite, and the perception of pain. Disruptions within the hindbrain's serotonin system have been implicated in a range of neuropsychiatric disorders (including depression and anxiety), neurodegenerative diseases (like Alzheimer’s and Parkinson’s), and various developmental abnormalities.

Hindbrain organoids represent cutting-edge three-dimensional in vitro models that capture the complexity of the developing human hindbrain, including those vital serotonergic neurons critical for emotional regulation, motor control, and diverse neuropsychiatric functions. Our Hindbrain Organoids Modeling Service utilizes advanced stem cell technology to generate hindbrain organoids tailored to specific patients or disease states, derived from human induced pluripotent stem cells (iPSCs). These sophisticated organoids are engineered to emulate the characteristics of caudal rhombomeres (r5–r8) and are enriched with functional serotonin (5-HT) neurons. This enables researchers to investigate neurodevelopmental processes, neurodegenerative diseases (such as Alzheimer’s), and neuropsychiatric symptoms (NPS) within a system that closely mirrors human biology.

By integrating well-established protocols validated in leading scientific studies, our service provides a robust and reliable platform for both drug screening initiatives and fundamental mechanistic investigations. Whether your research aims to explore the intricacies of serotonin signaling, model the agitation associated with Alzheimer’s disease, or test the efficacy of novel therapeutic interventions, our hindbrain organoids offer exceptional biological accuracy and the scalability needed for impactful research.

Key Advantages

1. High-Fidelity Modeling

• Generate organoids that accurately reflect the caudal hindbrain identity, featuring an enriched population of 5-HT neurons expressing key markers like TPH2, FEV, and GATA3, as rigorously validated through qPCR, flow cytometry, and immunocytochemistry.
• Replicate crucial aspects of human neurodevelopment, including the formation of neural tube-like structures, the presence of proliferative neural progenitors (NESTIN+, NKX2.2+), and the maturation of neurons (TUJ1+, MAP2+).

2. Functional Relevance

• Confirm the release of serotonin using ELISA assays and demonstrate physiological electrical activity, including both tonic and burst firing patterns, ensuring the model's functional authenticity.
• Effectively model drug responses, such as those to selective serotonin reuptake inhibitors (SSRIs) like escitalopram, and explore the impact of gut microbiome interactions, reflecting the clinical variability observed in treatment outcomes.

3. Patient-Specific Insights

• Utilize iPSCs derived from healthy individuals or patients diagnosed with Alzheimer’s disease, Parkinson’s disease, or various neuropsychiatric disorders to facilitate the study of inter-individual differences in disease progression and treatment response.

4. Scalability and Customization:

• Our optimized protocols consistently yield organoids with uniform size (1–2 mm diameter) and sustained viability for over 12 weeks, making them well-suited for high-throughput screening applications.
• Offer the flexibility to tailor these models with specific genetic modifications (e.g., introducing APOE or PSEN1 mutations) or through the implementation of co-culture systems (e.g., incorporating glia or vascular cells) to create more complex and nuanced disease models.

Our Comprehensive Brainstem Organoid Services

1. Hindbrain Organoid Generation

• Cell Sources: iPSCs derived from patient PBMCs, hPSCs (H9), or custom cell lines.
• Differentiation Protocol:

- Embryoid Body Formation
- Neural Precursor Induction
- Maturation

Fig.1 Generation of hindbrain organoids from iPSCs.^1,2

• QC Metrics: Size/circularity tracking, NPC markers (Nestin, NKX2.2), and 5-HT neuron markers (5-HT, TUJ1).

Fig.2 Hindbrain organoid stained with NPC markers Nestin (green) and NKX2.2 (in red).^1,3

Fig.3 Immunocytochemistry for the hindbrain organoids.^1,3

2. Drug Screening and Pharmacology

• SSRI Testing: Dose-response assays for escitalopram, sertraline, or novel compounds, measuring extracellular 5-HT levels (ELISA) and transcriptional changes (TPH2, SERT).
• Toxicity Studies: Assess metabolic activity and neurodegeneration (β-amyloid/tau seeding).

3. Disease Modeling

• Alzheimer’s Disease: Model NPS (agitation, depression) using organoids from AD patients with/without neuropsychiatric symptoms.
• Parkinson’s Disease: Investigate serotonin-dopamine crosstalk in motor and mood-related pathways.

4. Customized Solutions

• Multi-Omics Integration: Combine transcriptomics, proteomics, and electrophysiology for mechanistic insights.
• Co-Culture Systems: Incorporate microglia, astrocytes, or gut microbiota-derived metabolites to study neuro-immune/metabolic interactions.

Our Workflow

Transform Your Research with Human-Relevant Hindbrain Models

From basic neurodevelopment to clinical drug discovery, our Hindbrain Organoids Modeling Service bridges the gap between in vitro assays and human physiology. Contact us to design a study tailored to your scientific goals—whether unraveling serotonin pathways or accelerating patient-specific therapeutics.

References

1. Zivko, Cristina et al. “iPSC-derived hindbrain organoids to evaluate escitalopram oxalate treatment responses targeting neuropsychiatric symptoms in Alzheimer's disease.” Molecular psychiatry vol. 29,11 (2024): 3644-3652. doi:10.1038/s41380-024-02629-y
2. Distributed under Open Access License CC BY 4.0. The original image was modified.
3. Distributed under Open Access License CC BY 4.0 without modification.