Ganglion Organoid Modeling Service

The peripheral nervous system (PNS) comprises a complex network essential for bodily function, containing ganglia—clusters of specialized nerve cell bodies. These ganglia fall into two major functional categories: Sensory Ganglia, which process and relay information about our external and internal environment (touch, pain, temperature, proprioception, visceral status), and Autonomic Ganglia, which orchestrate involuntary functions like heart rate, digestion, and respiration via the sympathetic and parasympathetic systems.

Developing effective therapies for conditions involving the PNS—such as chronic pain, sensory neuropathies, gut-brain axis disorders, and autonomic dysfunction—requires models that accurately reflect human physiology. Creative Biolabs addresses this need by providing specialized Ganglion Organoid Modeling Services, utilizing advanced 3D organoid technology derived from human induced pluripotent stem cells (iPSCs) and directly reprogrammed fibroblasts.

Our Specialized Ganglion Organoid Models & Services

We offer tailored services focusing on distinct ganglion types, enabling targeted research into specific PNS functions and pathologies:

A. Sensory Ganglion Organoid Platform

This platform encompasses organoid models representing ganglia responsible for detecting and transmitting sensory inputs from the body surface and internal organs.

1. Dorsal Root Ganglion (DRG) Organoids:

Model System: We generate human iPSC-derived DRG organoids (hDRGOs) using refined, developmentally informed protocols. By incorporating key signaling factors identified through spatiotemporal transcriptomic analysis of human embryonic DRGs (e.g., precise modulation of WNT, FGF, RA, and neurotrophin pathways), our hDRGOs faithfully recapitulate critical aspects of human DRG formation in vitro.

Features: hDRGOs contain a rich diversity of physiologically relevant sensory neuron subtypes, including nociceptors, mechanoreceptors, and proprioceptors, alongside essential satellite glial cells and Schwann cells. These organoids express key markers (NTRK1/2/3, RUNX1/3, PIEZO2) and form functional neuronal networks. Notably, we can generate models containing unique human-enriched nociceptor subtypes (e.g., NTRK3+/DCC+ neurons) that exhibit characteristic responses to stimuli like capsaicin. Neurons within these organoids display mature features, including pseudo-unipolar morphology and expression of functional ion channels (SCN10A, SCN11A).

Services & Applications: Generation of hDRGOs from control or patient-derived iPSCs; comprehensive characterization (IHC/ICC, spatial transcriptomics, calcium imaging, electrophysiology); TF function studies (e.g., via shRNA knockdown); modeling somatic sensory modalities (touch, pain, temperature, proprioception); investigating neuropathic pain mechanisms; screening for novel analgesics and neuroprotective agents.

Fig.1 Generation and characterization of human DRG organoids.¹

2. Visceral Sensory Ganglion Organoids (VSGOs):

Model System: We provide human iPSC-derived VSGOs differentiated via carefully guided protocols mimicking epibranchial placode development, specifically modeling ganglia like the nodose ganglion involved in monitoring internal organ status.

Features: VSGOs contain VSNs expressing canonical markers (PHOX2B, ISL1) and a diverse array of functionally relevant receptors (GLP1R, CCKAR, HTR3A, TRPV1, NPY1R, SSTR2, PIEZO1) essential for detecting chemical and mechanical signals from the viscera. Single-cell transcriptomics confirms cellular heterogeneity aligned with native VSN populations. VSGO neurons exhibit appropriate functional responses to stimuli such as GLP1R agonists (liraglutide) and capsaicin.

Services & Applications: Generation of VSGOs from control or patient-derived iPSCs; modeling visceral sensation and pain; studying gut-brain axis communication, particularly when integrated into microfluidic 'Axis-on-a-Chip' systems with human colon organoids (HCOs); investigating VSN roles in metabolic disorders or neurodegenerative diseases (e.g., AD, exploring APOE4-dependent pathological protein propagation); drug screening targeting visceral afferents.

3. Induced Sensory Ganglion (iSG) Organoids:

Model System: Leveraging direct reprogramming technology, we generate self-organizing iSG organoids from human or mouse fibroblasts using defined transcription factors (Ascl1, Brn3a/b, Isl1 - 'ABI' factors). This approach bypasses progenitor stages, directly converting fibroblasts into networked sensory neurons.

Features: iSGs spontaneously form interconnected neuronal clusters containing heterogeneous sensory neuron populations expressing subtype markers (TrkA, TrkB, TrkC, NF200, Peripherin) and functional markers (VGLUT1/2, Synapsin). These neurons exhibit mature electrophysiological properties (single/multiple action potentials, synaptic activity) and respond appropriately to sensory stimuli (capsaicin, menthol). They demonstrate the capacity for peripheral innervation in vitro.

Services & Applications: Generation of iSGs from client-provided fibroblasts (control or patient-derived); an alternative platform for sensory neuron research, particularly useful when starting from easily accessible somatic cells; functional characterization; modeling hereditary sensory neuropathies (e.g., Friedreich's ataxia) or channelopathies using patient cells; studying peripheral innervation patterns in co-culture models (e.g., with skin cells).

B. Autonomic Ganglion Organoid Platform

This platform focuses on ganglia of the autonomic nervous system (ANS), which controls involuntary bodily functions through its sympathetic and parasympathetic branches.

1. Induced Autonomic Ganglion (iAG) Organoids:

Model System: We generate self-organizing iAG organoids by directly reprogramming mouse or human fibroblasts using a potent combination of transcription factors (Ascl1, Phox2a/b, Hand2 - 'APH' factors).

Features: iAGs form networked clusters containing diverse autonomic neuron subtypes, including cholinergic (ChAT+, VACHT+) and noradrenergic (TH+, DBH+) neurons, reflecting the heterogeneity of the ANS. These neurons express synaptic markers (Synapsin) and exhibit mature electrophysiological profiles (action potentials, tonic/phasic firing, neurotransmitter responses to ACh, dopamine, noradrenaline). Functionally, iAG neurons can innervate target cells, such as cardiomyocytes, and modulate their activity (e.g., beating rate) in vitro. Transcriptomic analysis confirms their identity and similarity to endogenous AG neurons.

Services & Applications: Generation of iAGs from control or patient fibroblasts; modeling autonomic neuropathies (e.g., AAG, diabetic autonomic neuropathy) or dysautonomia associated with neurodegenerative diseases (PD); studying sympathetic/parasympathetic development and function; functional testing via co-culture assays (e.g., iAG-cardiomyocyte); screening for drugs modulating autonomic activity.

Custom Autonomic Models: We also offer development of iPSC-derived autonomic ganglion models tailored to specific research needs.

Cross-Platform Capabilities & Advanced Applications

To maximize the utility of our ganglion organoid models, we offer a range of advanced analytical and application-focused services:

• Comprehensive Characterization: Detailed molecular profiling via sc/snRNA-seq; high-resolution imaging with multiplex IHC/ICC for protein localization; spatial transcriptomics; functional validation using live calcium imaging (GCaMP or dyes) and patch-clamp electrophysiology.
• Advanced Disease Modeling: Utilization of patient-derived iPSCs or fibroblasts; targeted gene editing (CRISPR/Cas9) or knockdown (shRNA) to investigate gene function; development of complex co-culture models incorporating immune cells or specific target tissues.
• Drug Discovery Platform: Scalable organoid models suitable for medium-to-high-throughput screening (HTS) of compound libraries; detailed mechanism-of-action studies; assessment of drug efficacy and neurotoxicity on human neuronal subtypes.
• Circuit & Pathway Modeling:
  • Assembloids: Engineering multi-organoid systems, such as the 4-part Human Ascending Somatosensory Assembloid (hASA: hSeO-hdSpO-hDiO-hCO), to model long-tract pathways ex vivo. Enabling investigation of synaptic transmission, network synchrony, pathway development, and circuit-level effects of genetic variants (e.g., SCN9A) or drugs.
  • Axis-on-a-Chip: Integrating VSGOs with HCOs, or iAGs with relevant target organoids (e.g., cardiac), on microfluidic devices to study bidirectional neuro-organ communication, barrier function, and disease propagation in controlled microenvironments.
  • Innervation Models: Custom co-culture systems to model innervation of specific peripheral targets (skin, muscle, heart tissue) by sensory or autonomic neurons.

Applications

Our Ganglion Organoid Modeling Services empower research across multiple domains:

• Pain Research: Nociception, neuropathic & inflammatory pain, channelopathies, analgesic discovery.
• Sensory Neuropathies: Modeling hereditary, metabolic (diabetic), or toxic neuropathies.
• Neurodegenerative Diseases: Investigating PNS involvement, autonomic failure, and gut-brain contributions (AD, PD).
• Gut-Brain Axis Research: Visceral sensation, microbiome-nerve interactions, metabolic signaling, inflammatory bowel disease.
• Autonomic Nervous System Disorders: Dysautonomia, cardiovascular regulation, Hirschsprung's disease models.
• Developmental Neurobiology: Studying human PNS specification, migration, differentiation, and synaptogenesis.
• Drug Discovery & Toxicology: Target identification/validation, compound screening, predictive toxicology.

Our Workflow

Explore the intricacies of the human peripheral nervous system using Creative Biolabs' specialized ganglion organoid and assembloid models. Our advanced platforms provide physiologically relevant, human-centric tools to accelerate your research and therapeutic development efforts.

Reference

1. Lu, Tian et al. "Decoding transcriptional identity in developing human sensory neurons and organoid modeling." Cell vol. 187,26 (2024): 7374-7393.e28. doi:10.1016/j.cell.2024.10.023. Distributed under Open Access License CC BY 4.0. The original image was modified.