3D Organoids

CNS-3D Organoids are derived from donor-induced pluripotent stem cells (iPSCs) differentiated into neural progenitor cells (NPCs). The resulting structures grow to 500–800 µm in diameter, exhibit spontaneous, electrically driven network activity, and display transcriptomic profiles consistent with human cortical development

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Cell Types

CNS-3D Organoids are comprised of ~50% neurons and ~50% astrocytes which result from the co-differentiation of a single neural progenitor cell suspension. Of this neuron population, 95% are glutamatergic (excitatory) neurons and ~5% are GABAergic (inhibitory) neurons. SnRNAseq analysis confirms that CNS-3D Organoids exhibit a mature biological profile, characterized by a large supporting astrocyte population and a small progenitor cell population.

Reproducibility

Consistent and stable organoid pharmacology enables reliable, reproducible results from the first experiment, supports longitudinal (weeks-long) treatment paradigms, and maintains data quality across a validated 4-week assay window.

Quality Control

Each shipment includes a certificate of analysis detailing organoid quality, including brightfield imaging to confirm size consistency (500–800 µm, <5% CV, 98% presence), neuron-to-astrocyte ratios via ICC, sterility (mycoplasma, bacterial, fungal), and assay performance metrics such as EC/IC₅₀and robust Z’ values for FLIPR screening.

Assays

FLIPR Functional Modulation

CNS-3D Organoids exhibit spontaneous functional activity quantifiable via high-throughput FLIPR using a calcium-sensitive fluorophore. Consistent oscillatory waveforms across replicates enable reliable neurotoxicity and neuromodulation screening with as few as four replicates per condition.

Cell Viability Assessment

Cell viability assays like CellTiter-Glo and LDH-Glo can be run alone to reveal dose-dependent effects on organoid health or multiplexed with FLIPR to differentiate neuromodulation from toxicity-driven functional changes.

3D High-content Imaging

High-resolution imaging with custom analysis algorithms enables precise quantification of treatment-induced morphological changes in specific brain cell types at cellular, subcellular, and projection levels.

Multiplexed Sample Collection

Post-treatment CNS-3D Organoids and media can be frozen to preserve RNA, proteins, and secreted factors for downstream analyses, including transcriptomics, proteomics, and ELISA-based biomarker assays.

Additional Assays

CNS-3D Organoids support standard fluorescent, luminescent, and colorimetric assays, including JC-10 for mitochondrial health, Live/Dead imaging for cell-type-specific toxicity, cytokine profiling, and enzymatic activity (e.g., AChE).

Use Case 01

Neurotoxicity

A study published in ALTEX found that CNS-3D Organoids were 7.41 times more accurate than traditional animal models at predicting clinical neurotoxicity because they contain multiple iPSC-derived neural cell types that capture the complexity of human brain tissue. Unlike basic cell cultures, CNS-3D Organoids measure functional electrical activity, enabling detection of early changes in neural network behavior that may indicate toxicity before structural damage occurs.

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Dose response data of 8 representative compounds with various degrees of toxicity as assessed by potency of neuromodulation (peak count) relative to clinically relevant concentrations (Cmax).

Use Case 02

Disease Modeling

CNS-3D Organoids have been used to model two rare, X-linked neurodevelopmental disorders – Rett Syndrome (RTT) and CDKL5 Deficiency Disorder (CDD). Though these two distinct disorders exhibit overlapping phenotypic features including cognitive deficits, developmental delays, and seizures, when modeled using CNS-3D Organoids we observe striking differences in each disease model in terms of their spontaneous calcium bursting activity profiles.

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CNS-3D organoids modeling two distinct neurodevelopmental disorders Rett Syndrome (orange) and CDKL5 Deficiency Disorder (teal) display unique functional phenotypes and can be grown in 384-well plate formats.

Use Case 03

Combined Safety & Efficacy Assessment

CNS-3D Organoids derived from healthy control and CDKL5 Deficiency Disorder (CDD) patients were used to identify novel disease-modifying therapeutics. Leveraging a custom brightfield imaging-based predicted toxicity algorithm, we simultaneously assessed compound safety and efficacy—enabling selection of candidates that selectively rescued the CDD FLIPR phenotype without impacting viability or healthy control organoid function.

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Dose response data demonstrating disease-selective rescue without impacts to cell viability (predicted CTG). Compound restores aberrant CDD organoid function (teal) to Control levels (white).

Use Case 04

Neuromodulator Profiling

Using FLIPR and CTG assays, a study performed in 2019 (Sirenko, 2019 Toxicology) showed that the organoids could detect subtle, dose-dependent effects of known pharmaceuticals, environmental pesticides, industrial compounds and more on neuronal network activity; identifying functional changes prior to overt cytotoxicity. The results highlight the organoids utility for early-stage safety screening, profiling the effects of compounds on neuromodulator-regulated network dynamics and mechanism-of-action studies in CNS drug development.

View Poster

CNS-3D organoids treated with nine representative compounds cause unique functional phenotypes to appear in fluorescence over time calcium imaging experiments.

Engineered brain organoids: 7.4x more predictive of human clinical outcomes.

A Complex in vitro Model

Neurotoxicity

Disease Modeling

Combined Safety & Efficacy Assessment

Neuromodulator Profiling

Products & Services

CNS-3D Technology Resources