In the dynamic landscape of neuroscience, the mission to transform the future of neurological therapeutics requires tools as powerful as the innovative scientific visions driving the research. Human pluripotent stem cells (hPSCs) have emerged as a promising tool for biotech companies. Inspired by knowledge, innovation, and quality, STEMCELL Technologies’ tools and services are helping scientists accelerate the pace of discovery.
hPSC-derived neural models are revolutionizing the efficiency of the drug development process for biotech companies by providing a more accurate and relevant alternative to traditional animal and simpler in vitro models. hPSCs are self-renewing, multipotent cells and so can be differentiated into neural cells. These hPSC-derived neural models, including neurons, glia, organoids, and co-cultures, stand out for their ability to effectively recapitulate complex human neural physiology, providing an essential platform for understanding the complexities of neurological diseases and developing effective treatments.
Drug developers aim to make faster and more reliable decision-making early in the therapeutic development process to mitigate the high costs associated with late-stage clinical trial failures due to discrepancies in drug effects between non-human models and actual human outcomes. The physiological relevance of these advanced models enables drug developers to make decisions using more predictive data. hPSC-derived neural models also address the high drug attrition rate by supporting the creation of refined disease models. hPSC models allow for detailed investigations into a treatment’s toxicity and physiological interactions within a context that closely mimics human neural tissue, using either patient-derived or genetically engineered cells to replicate human cellular diversity. Furthermore, these models provide comprehensive data enabling informed decision-making throughout drug development. This significantly enhances the likelihood of clinical success by identifying therapeutic effects early on and providing the confidence needed to advance therapeutic candidates into clinical trials. Ultimately, integrating hPSC-derived neural models into drug development pipelines offers the opportunity for more efficient, cost-effective solutions that are more predictive of human outcomes, thereby reducing the likelihood of late-stage clinical failures.
With fewer than 10% of the compounds that enter Phase I clinical trials being approved for clinical use and more than 75% of these failures being due to efficacy and/or safety issues (1), more predictive models are of paramount importance. From target discovery to preclinical testing, hPSC models inform decisions rapidly, reliably, and predictively.
hPSC-based neural workflows provide a robust tool to leverage patient-specific disease models, allowing for more accurate drug efficacy predictions, especially when studying rare pathologies. For example, researchers from the University of Manitoba developed a clinical trial pre-screening platform to assess therapeutic effectiveness for patients with ultra-rare diseases using the personalized potential of induced pluripotent stem cell (iPSC) models.2 With human brain tissue being hard to come by, the ability to generate patient-specific differentiated cells from iPSCs offers unique potential. Using a controlled, patient-specific disease model context to evaluate drug candidates enables more informed decisions about lead selection.
To create a robust platform for assessing the efficacy and toxicity of the drug panel of interest, developing a high-quality iPSC-derived neural model was crucial for this research. The researchers from the University of Manitoba used STEMdiff™ Forebrain Neuron Differentiation Kit for highly pure and efficient neural differentiation (Figure 1) of patient-derived iPSCs for enhanced reliability and reproducibility. The team also used STEMdiff™ Forebrain Neuron Maturation Kit, which supports long-term neural cell culture maintenance and promotes neuronal activity, providing physiologically relevant results.
Figure 1. Highly efficient differentiation of neural progenitor cells to neurons using STEMdiff™ forebrain neuron differentiation and maturation kits.
(A) NPCs generated from STiPS-R038 hPSCs in mTeSR™1 using STEMdiff™ SMADi Neural Induction Kit embryoid body protocol were differentiated and matured to cortical neurons using STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and STEMdiff™ Forebrain Neuron Maturation Kit for 14 days. The resulting cultures contain a highly pure population of (B) class III β-tubulin-positive neurons (green) with less than 10% GFAP-positive astrocytes (not shown). (C) The generated neurons are also positive for FOXG1 expression (red), indicating a forebrain-type identity. (D) Nuclei are labeled with Hoechst (blue). NPCs = neural progenitor cells; hPSC = human pluripotent stem cell.
Researchers from the Drug Safety Research & Evaluation team at Takeda Pharmaceuticals have investigated innovative methods to predict neurotoxicity during the lead optimization process. The team evaluated applying an iPSC-derived 3D neural model as a high-throughput predictive assay. They determined that the neural spheroids maintained high specificity in identifying human neurotoxicity concerns.3 Using iPSC-derived 3D models for lead optimization enables a reduction in the number of animals used to narrow down candidate compounds and a more relevant model with human-specific cellular biology and genetics when assessing neurotoxicity.
The 3D neural spheroids were cultured in a physiologically relevant environment, using BrainPhys™ Neuronal Medium. Choosing a medium that supports long-term culture and enhances neuronal function is critical for accurately assessing neurotoxicity in drug candidates and achieving predictive results.
Specialized 3D hPSC-derived models can recapitulate complex human-specific biology in ways that alternative models cannot. hPSC-derived choroid plexus organoids develop physiological cystic structures that mimic the complexity of the human blood-cerebrospinal fluid (CSF) barrier.4 This type of advanced model allows for a deeper understanding of the permeability of neuroactive drugs across this blood-CSF barrier for more relevant insights.
When using organoid models for performing drug screening, organoid reproducibility is critical to achieve reliable and translatable results.STEMdiff™ Choroid Plexus Organoid Differentiation Kit and STEMdiff™ Choroid Plexus Organoid Maturation Kit enable the generation of reproducible organoids with CSF-like fluid cysts, which can be maintained long-term for screening applications (Figure 2).
Figure 2. STEMdiff™ Choroid Plexus Organoid Differentiation Kit Supports the Generation of Reproducible Organoids
Day 50 choroid plexus organoids generated with STEMdiff™ Choroid Plexus Organoid Differentiation Kit in a 6-well plate.
When high-quality results are at the core of what you do, consistent hPSC differentiation is crucial. Without standardized hPSC culture conditions, even the most detailed and rigorously followed stem cell differentiation protocols may still lead to inconsistent differentiation.5,6 STEMdiff™ is a line of culture medium kits specifically optimized for hPSC differentiation. Whether you’re looking to produce 2D neural cell types or 3D organoid models, there are standardized kits to reproducibly and efficiently differentiate embryonic stem and induced pluripotent stem cell lines. These kits offer an easy-to-use workflow that’s accessible regardless of your level of hPSC culture experience, while still maintaining the benefits of consistency and physiological relevance.
One consideration with hPSC-based models is that these cells are inherently immature, a potential limitation when developing a predictive drug discovery model. To overcome this and promote maturity in hPSC-derived neural systems, assessing physiologically relevant neural activity is important. With long-term culture, stronger and more coordinated network activity develops in hPSC-derived neural cultures, reflecting a more mature state.BrainPhys™ Neuronal Medium is a neurophysiological basal medium that promotes neuronal function and synaptic activity, enhancing the physiological relevance of hPSC-based neural models.7 Neural STEMdiff™ kits also integrate these benefits with BrainPhys™-based maturation media.
Figure 3. hPSC-derived neurons generated in BrainPhys™ neuronal medium express markers of neuronal maturity after 14 and 44 days of differentiation.
Figure 4. hPSC-derived neurons matured in BrainPhys™ neuronal medium show improved excitatory and inhibitory synaptic activity.
NPCs were generated from H9 cells using STEMdiff™ Neural Induction Medium in an embryoid body-based protocol. Next, NPCs were cultured in (A,C) BrainPhys™ Neuronal Medium, supplemented with 2% NeuroCult™ SM1 Supplement, 1% N2 Supplement-A, 20 ng/mL GDNF, 20 ng/mL BDNF, 1 mM db-cAMP, and 200 nM ascorbic acid to initiate neuronal differentiation, or (B,D) DMEM/F-12 under the same supplementation conditions. After 14 and 44 days of differentiation and maturation, neurons express the synaptic marker Synapsin 1 (green) and the mature neuronal marker MAP2 (red). In this example, neurons matured in BrainPhys™ Neuronal Medium show increased Synapsin 1 staining. Scale bar= 100 µm.
Even without hPSC culture experience, adopting hPSC-based models is becoming increasingly accessible. Cryopreserved cell products at various stages of neural differentiation make getting started quickly simple and can save you time, skipping stages of the hPSC workflow. Start with a high-quality iPSC cell line like SCTi003-A (female) or SCTi004-A (male) for optimal performance and reproducibility. Use a highly characterized neural progenitor intermediate like Human iPSC-Derived Neural Progenitor Cells for a confident start to your neural workflow, which can be differentiated into various neural cell types (Figure 5). Otherwise, jump right into experimentation with a highly pure population of Human iPSC-Derived Forebrain Neuron Precursor Cells. Alternatively, for a 3D model, choose Human iPSC-Derived Midbrain Organoids and culture these with STEMdiff Neural Organoid Maintenance Kit for optimal performance.
Figure 5. Human iPSC-derived neural progenitor cells can effectively differentiate into forebrain neurons, midbrain neurons, and astrocytes.
Human iPSC-Derived Neural Progenitor Cells generated from SCTi003-A iPSCs were thawed, established in culture, and fixed for immunocytochemistry. (A) The NPCs express neural progenitor markers SOX1 (red) and PAX6 (green). (B) NPCs cultured with the STEMdiff™ Forebrain Neuron Kit produce forebrain neuron cell populations expressing neuronal identity marker βIII-TUB (magenta). (C) NPCs cultured with the STEMdiff™ Midbrain Neuron Kit produce midbrain neuron cell populations expressing neuronal identity marker βIII-TUB (red) and dopaminergic neuron marker TH (green). (D) NPCs cultured with the STEMdiff™ Astrocyte Kit produce astrocyte populations expressing astrocyte marker S100β (green) and GFAP (red). iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells.
For more information about STEMCELL's neural portfolio , educational resources ,or if you would like to learn more about how we can support your neuroscience research,please contact us attechsupport@stemcell.com.
References
1. Sun D et al. (2022) Why 90% of clinical drug development fails and how to improve it? Acta Pharm Sin B 12(7):3049–62
2. Sequiera GL et al. (2022) Development of iPSC-based clinical trial selection platform for patients with ultrarare diseases. Sci Adv 8(14):eabl4370
3. Wang Q et al. (2022) Assessment of a 3D neural spheroid model to detect pharmaceutical-induced neurotoxicity. ALTEX 39(4)560–82
4. Chew LH et al. (2024) Methods to extract and analyze fluid from human pluripotent stem cell-derived choroid plexus organoids. Front Mol Neurosci 16:1243499
5. D’Amour KA et al. (2005) Efficient differentiation of human embryonic stem cells to definitive endoderm. Nat Biotech 23(12):1534–41
6. Kattman SJ et al. (2011) Stage-specific optimization of activin/nodal and bmp signaling promotes cardiac differentiation of mouse and human pluripotent stem cell lines. Cell Stem Cell 8(2): 228–40
7. Bardy C et al. (2015) Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro. PNAS 112(20):E2725–E2734