| Overview |
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The most common neurological diseases include stroke, traumatic brain injury, drug addiction, Alzheimer's disease, Parkinson's disease, anxiety, depression and etc. This group of diseases constitutes one of the biggest social, healthy and financial problems for human |
being. To fight against these diseases is currently an extensive focus in the fields. |
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From end to end, we can support your neuroscience research through our experience/expertise from both pharmaceutical industry and academia. HDB offers comprehensive discovery platform in both In-vitro neuroscience and In-vivo neurological disease models. Find out how to partner with us to accelerate the drug discovery for the |
treatment of these devastating disorders. |
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| In-Vitro Neurosciences Platform Overview and Selected Assay List |
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Assay and Technique |
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Target based biochemical and cellular assays |
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Virus based gene delivery and shRNA/CRISPR Knock out, knock down, |
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overexpression, knockin |
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Inflammation markers (high content imaging, RT-PCR, ELISA, IHC, etc.) |
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Microglia, astrocytes biology and functional assays |
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Cell viability/Apoptosis; Neurite outgrowth |
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Neuroprotective assessment in neurotoxcin induced cytotoxicity assay using |
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neuronal cell line, rodent primary neuron, or rodent primary glia |
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Protein expression, production and phosphorylation (MSD, Western blot, ELISA, JAZZ) |
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IHC and pathology |
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3D and organoids |
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iPD cells differentiation and characterization |
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Cell type |
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Primary neurons: Dorsal root ganglion neurons, Cortical neurons, Hippocampus |
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neurons, norepinephrine (NE) neurons, etc. |
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Neuronal cell lines: PC12, GT1-7, etc. |
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Primary pituitary cells |
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Primary microglia and astrocyte |
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Human iPS cells |
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Target based biochemical and cellular assays |
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AChR cluster formation |
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Inflammation markers (high content imaging assay, RT-PCR, ELISA, IHC, etc.) |
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Microglia, astrocytes biology and functional assays |
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NSCs Differentiation Assay |
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Cell viability/Apoptosis; Neurite outgrowth |
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Neuron Damage Assay |
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3D and organoids |
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CNS 3D organelle (iPSC base) characterization |
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Neuroprotective assessment in neurotoxin induced cytotoxicity assay using |
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neuronal cell line, rodent primary neuron, or rodent primary glia |
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Protein expression, production and phosphorylation (MSD, Western blot, ELISA, |
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JAZZ) |
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| In-vitro Assay Showcase |
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| Assay #1: α-Synuclein Degrader Compound Screen using HTRF Assay |
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| Endogenous α-Synuclein (α-Syn) HTRF evaluation was validated in SH-SY5Y (R2›0.95), and batches of a-Syn degrader compounds have been screened using this validated HTRF assay with in parallel cytotoxicity assessment by CTG. |
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| Assay #2: CNS-related for Cellular Kinase Assay |
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| 1. Successful developed kinase assay from WB format to AlphaLISA format suited to HTS |
| 2. Screened ~60,000 compounds with hit rate of 1.66% |
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| Assay #3: CNS-related GPCR Binding and Functional Assays |
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| Comprehensive in-vitro assay platform for GPCR targets |
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Cover stable cell line generation (›100 GPCR cells collection in |
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house), assay development, compound screening and profiling, |
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HTS |
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Multiple formats and technologies available |
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HTRF IP-1, cAMP |
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FLIPR |
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Radioligand binding |
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Reporter assays (CRE/reporter, NFAT/reporter) |
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| Assay #4: Dopamine and Glutamate Receptor Assay |
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| NMDA: N-methyl-D-aspartate (NMDA) receptor binding assays using membrane preparation from rat brain and purified proteins |
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| Assay #5: PP1 Binding Activity (FP Assay) |
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| Principle of PP1 FP Assay: |
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Protein phosphatase 1 (PP1) belongs to protein serine/threonine phosphatases family |
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PP1 plays an important role in neuronal functions |
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FP assay be designed to test compounds binding to PP1 |
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| Assay procedures |
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Prepare test compounds in 384-well LDV plate |
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Add 10ul FITC labeled peptide to assay plate |
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Add 10ul protein PP1 to assay plate |
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Incubate for 30min at room temperature |
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Read FITC FP on EnVision |
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| Assay #6: T-type Calcium Channel Assay in Rat DRG Neurons |
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| Panel A. |
| Sample traces from dose-response curve (DRC) of test compound using use-dependent pulse protocol (shown in the inlet). |
| Panel B. |
| DRC and calculated IC50 of test compound. Inhibition % is calculated as 1-(Icpd-Ivehicle)/Ivehicle*100% |
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| Assay #7: Aβ induced Phagocytosis and inflammation |
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Metal ions imbalance, a well-established pathologic feature of alzheimer’s disease |
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(AD), ultimately results in the deposition of amyloid-β peptide (Aβ) proteins and |
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Aβ-induced neurotoxicity. |
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One pathologic mechanism has been proposed that oligomeric Aβ oligomers can induce |
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microglia to secrete pro-inflammatory cytokines that lead to a reduction in the |
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expression of Aβ-binding scavenger receptors on microglia and eventual neurotoxicity. |
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Neurotoxicity from Aβ oligomers efficaciously restrained microglia’s phagocytosis and |
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prevented Aβ clearance. |
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| Assay #8: Aβ induced Phagocytosis and inflammation in Mice Primary Microglia or Astrocyte |
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| Application: Compound evaluation for Aβ induced phagocytosis and cytokine in mice primary microglia and astrocyte. |
| Highlights: HCS imaging on phagocytosis and qPCR detection for cytokine level |
| Data Examples: Positive compounds showed dose-dependent enhancement of Aβ phagocytosis and repression of IL-1β in mice primary microglia and astrocyte |
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| Assay #9: LPS induced inflammation in Mice Primary Microglia |
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| To assess compound effect on cytokine IL-1β secretion in mice primary microglia. |
| Cytokine IL-1β secretion by primary microglia was evaluated by ELISA. Positive compound showed dose-dependent repression of IL-1β in mice primary microglia. |
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| Assay #10: Tau Degradation Assay |
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| Purpose: To identify inhibitors for Tau fibrillization in mutant Tau overexpressed cell. |
| Description: Tau tangles as one of the pathological hallmark for Alzheimer’s disease is a potential therapeutic focus. Cells are subjected to the pre-formed fibril Tau (PFF) which will trigger the Tau phosphorylation and fibril Tau formation in the cells. The image was |
| captured by PE Operetta CLS using 20x water objective. |
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| Representative Images： |
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| Assay #11: Neurite Outgrowth Assay – HCS/PC12 |
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| Application: To quantify neurite outgrowth. |
| Highlights: HCS Imaging, Neurite Outgrowth Analysis |
| Data Example: PC12 cells were cultured and incubated with NGF in 96-well plate for 7 days. |
| NGF could promote neurite outgrowth with EC50 of 21.53 ng/ml in PC12 cells. |
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| Assay #12: Neurite Outgrowth Assay – HCS/hiPSC derived DA neurons |
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| Application: To quantify neurite. |
| Highlights: HCS Imaging, Neurite Outgrowth Analysis |
| Data Example: Midbrain dopaminergic (miDA) neurons derived from iPSC cells were grown in 384-well plates and stained with Calcein AM after 3, 5, and 15 days of in-vitro maturation. Neurite tracing and quantification (n = 6) shows increase of neurite length in |
| miDA neurons during in-vitro maturation. |
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| Assay #13: Neurite Outgrowth Assay – HCS/NSCs |
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| Application: To quantify neurite outgrowth. |
| Highlights: HCS Imaging, Neurite Outgrowth Analysis |
| Data Example: NGF and cAMP could promote neurite outgrowth in primary cultured rat |
| neural stem cells (NSCs). |
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| Assay #14: NSCs Differentiation Assay – HCS/NSCs |
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| Application: To quantify the differentiation degree of primary cultured rat neural stem |
| cells (NSCs). |
| Highlights: HCS Imaging, Differentiation Model, Multichannel Record |
| Data Example: Compared with vehicle control, the forskolin group showed increased |
| neuron and astrocyte cell number proportions. |
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| Assay #15: Various Assays in Primary Hippocampal Neurons |
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| Applicable disease models/assays：Excitotoxicity (glutamate, kainate), Neurodegeneration (Aβ or Tau), Neurite growth assay and others |
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| Assay #16: Various Assays in Primary Cortical Neurons |
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| Applicable disease models/assay: Excitotoxicity (glutamate, kainate), Neurodegeneration |
| (Aβ or Tau), Neurite growth assay, Drug induced gene expression |
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| Assay #17: Neurotoxicity Assay – MEA/Primary mice hippocampus neurons |
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| Evaluate drug induced neuron damage by detecting the electroneurographic signals using Maestro EDGE MEA. Hippocampus neurons were isolated from neonatal mice (C57, 1d) and cultured in CytoView MEA plate up to steady states. 10 days later neurons were incubated with Aβ for 72 hours. Then the Aβ was washed and neurons were cultured for another 72 hours. The spikes and synchrony index showed that 50μM Aβ induced hippocampus neurons damage with continuous 72 hours incubation. And the neurons |
| viability was recovered after Aβ washed. |
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| Drug induced neuron damage evaluation using MEA. |
| A. A representative movie of electric signals detected by 16 electrodes in one well. |
| B & C. The spikes and synchrony index showed that 50μM Aβ induced hippocampus neurons damage with continuous 72 hours incubation. And the neurons viability was |
| recovered after Aβ washed. |
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| Assay #18: Neurotoxicity – HCS/hiPSC derived DA neurons |
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| Evaluate drug induced neurotoxicity in dopaminergic neuron using HCS imaging system. |
| Midbrain dopaminergic (miDA) neurons derived from human iPSC cells were cultured in 96-well plate for 5 days. 6-OHDA was added into the plate and cultured for 24 hours. Imaged and analyzed in high-content imaging system to |
| quantify neurotoxicity induced by 6-OHDA. |
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| Assay #19: Brain Organoids from iPSC |
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| Brain organoids from human iPSC for compounds evaluation. |
| A brain organoid platform has been established using human iPS cells from normal individuals or patients. These brain organoids have been characterized with a panel of |
| markers for brain cells, including neurons, glia as well as neural progenitor cells. |
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| Characterization of Brain Organoids |
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| Neuroscience In-vivo Disease Models |
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| By combining strong model developing capabilities and over 15 years of experience, HDB’s in-vivo pharmacology platform provides extensive neurological disease models with comprehensive biomarkers/pathology capabilities to accelerate your drug discovery and development. Our broad neuroscience model collection includes ready-to-go models in Alzheimer's diseases, Parkinson's diseases, neuro-degenerative diseases, neuro-inflammation, depression, anxiety, pain, drug addiction and many other therapeutic areas. |
| Our experts also offer customized models to meet your specific R&D needs. |
| Find out more >>> |
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