In Vitro Neuroscience Assays

Assessing microglial activation is an essential yet challenging aspect of truly translational CNS research. Using purified cultures from adult mice, new translational microglial activation assays can effectively recapitulate microglial behavior in vivo. Below are the microglial assays offered...

• Adult Murine Microglia Purification & Stimulation: These assays can be used to both probe ex vivo microglial activation status from healthy or diseased mice and assess whether immunomodulating therapeutics can dampen or enhance microglial activation in vitro.

• Microglial Activation Phagocytosis Assays: Adult murine microglia cultured under serum-free conditions efficiently phagocytose bio-particles and their phagocytic function can be either enhanced or suppressed depending on the mechanistic intervention. In vitro screening utilizing adult microglia derived from healthy or disease model tissue can provide information on mode of action and impact on cell function of therapeutics targeting microglial cells.

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CNS cells derived from human iPSCs have recently emerged as a straightforward, efficient, and replicable in vitro system for assessing aspects of human neuronal function such as neurite outgrowth. These cultures can be derived from healthy or patient-derived iPSCs. They can be studied in monoculture or co-culture to explore single cell mechanisms as well as the influence of neighboring cell types and signaling interplay. This iPSC technology is ideal for mechanistic studies and disease modelling in human cells.

Methodology to asses label-free neurite dynamics

Human iPSCs are terminally differentiated into neurons using a specific neuronal differentiation protocol. The effect of a therapeutic compound or inflammatory stimuli on cell viability and neurite outgrowth dynamic are assessed by NeuroTracker software.

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Charles River scientists have pioneered ion channel screening and research for nearly 20 years, creating more than 120 functionally validated assays and cell lines. We offer a variety of cutting edge technologies for ion channel screening, profiling and mechanism of action (MOA) studies, supporting our partners’ quests for the discovery of novel neuroscience ion channel therapeutics.

• Neurodegeneration/Stroke: Our Neurodegeneration/Stroke Channel Panel® includes ion channels which have been linked to disorders of the central and peripheral nervous systems. Voltage-gated sodium and potassium channels are potential neuroprotective therapeutic targets in ischemic stroke.

• Pain/Inflammation: Obtain rich data and scientific insights often not found at inhouse labs by accessing experienced Charles River scientists in the identification and characterization of a target compound for pain. Advanced ion channel screening quickly and efficiently identifies and characterizes a compound that modulates the activity of voltage-gated sodium, calcium, or potassium channels, or ligand-gated ion channels.

• Psychiatric Disorder: Our Psychiatric Disorder Channel Panel® includes neurotransmitter receptor channels that are important in anxiety and schizophrenia.Our Psychiatric Disorder Channel Panel® includes neurotransmitter receptor channels that are important in anxiety (GABA) and schizophrenia (NMDA and nAChRα7). Voltage-gated potassium channels (KCNQ) that regulate excitability and are potential therapeutic targets for anxiety and schizophrenia also are included.

• Seizure/Convulsion: Our Seizure/Convulsion Channel Panel® includes ion channels expressed in the central nervous system (CNS) that have been linked to inherited forms of epilepsy.Gain-of-function mutations in excitatory channels have been shown to cause persistent, depolarizing currents leading to hyperexcitability that facilitates epileptic seizures and convulsions

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We offer an extensive portfolio of in vitro models and in vitro assays to study Neurodegeneration. Disease-relevant models include immortalized neuroblastoma cells, induced pluripotent stem cells, and primary rodent neurons that can be modified using gene editing technologies (RNAi/CRISPR). These models are typically differentiated into neuronal precursor cell lines such as dopaminergic neurons. Several phenotypic screens are available using high content imaging-based methods to measure specific readouts.

• Alpha-synuclein aggregation: Developed in collaboration with The Michael J. Fox Foundation’s Parkinson’s Disease Research Tools Consortium, this Parkinson’s disease in vitro assay measures alpha-synuclein aggregation in a PD cell model using an aggregate selective anti alpha-synuclein antibody (MJFR14).

• TOM20 Loss Mitophagy Assay: Targeting the mitophagic regulators has gained momentum as a therapy based on their link to the gene mutations Parkin and PINK1. This Parkinson’s disease in vitro assay measures the loss of the structural mitochondrial marker TOM20 (TOMM20).

• MPP+ Neuronal Cell Death Assays: N-Methyl-4-phenylpyridinium Iodide (MPP+) is a neuron selective toxin that induces Parkinsonism in animal models. An active (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) MPTP metabolite, MPP+ has been shown to interfere with oxidative phosphorylation and induce mitochondrial dysfunction, resulting in depleted mitochondrial ATP production, and ultimately, the death of neurons. Using high content imaging, we can monitor cell death by measuring levels of propidium iodide that selectively accumulates in the dead neurons.

• Huntingtin (HTT) Detection Assays: CRL has a portfolio of Huntingtin (HTT) detection assays to support Huntington’s Disease (HD) drug discovery and development. Assays specific for different HTT protein species are available on the Mesoscale platform and a subset on Quanterix platform. Assays are well-validated in HD and non-HD animal models and include mutant HTT, aggregated HTT and rodent/NHP endogenous HTT. 

• Alzheimer's Disease Assays: The proper combination of Alzheimer’s disease in vitro assays can deliver critical information delivering insights that serve as better predictors for the transition to in vivo studies. Improve your in vivo success by integrating translational cell-based assays from simple cell proliferation and viability assays to multicellular biochemical and functional assays for high content phenotypic screening using our unique access to Alzheimer’s Disease relevant human primary cells from tissue, blood and differentiated stem cells into your AD discovery program 

• Amyotrophic Lateral Sclerosis Assays: Charles River has established an in vitro model from cultures of human induced pluripotent stem cells (hiPSCs) from patient cell lines harboring a mutation in one of the 4 genes which account for the majority of familial ALS. We have optimized a robust motor neuron differentiation protocol that is amenable to high-throughput screening. Using this protocol, approximately 80-85% of the differentiated neurons express mature motor neuron markers. This physiologically relevant cell system combined with a scalable differentiation protocol, allows for high-throughput screening of small molecule libraries or functional genomics-type approaches (RNAi / CRISPR-Cas9), using high content imaging or biomarker readouts.

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