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NeuroProof

Rostock, DE

NeuroProof as a contract research organization is a global leader and preferred partner in the field of microelectrode array (MEA)-based phenotypic screening of primary cultures. NeuroProof offers screening services and state-of-the-art mathematical expertise for the phenotypic analysis of test agents on neuronal network activity. The services comprise the growing of primary neuronal cultures on MEA chips, medium throughput screening of test agents, high quality recording of spontaneous and compound-induced network activity, state-of-the-art mathematical analyses, and sophisticated pattern recognition methods. The unique platform is well suited to identify and characterize effects of test agents on neuronal network activity and brain cell functioning.

Our company is specialized in the measurement of neuronal network activity via microelectrode arrays (MEA)-neurochips. We have extensive experience with a range of... Show more »

NeuroProof as a contract research organization is a global leader and preferred partner in the field of microelectrode array (MEA)-based phenotypic screening of primary cultures. NeuroProof offers screening services and state-of-the-art mathematical expertise for the phenotypic analysis of test agents on neuronal network activity. The services comprise the growing of primary neuronal cultures on MEA chips, medium throughput screening of test agents, high quality recording of spontaneous and compound-induced network activity, state-of-the-art mathematical analyses, and sophisticated pattern recognition methods. The unique platform is well suited to identify and characterize effects of test agents on neuronal network activity and brain cell functioning.

Our company is specialized in the measurement of neuronal network activity via microelectrode arrays (MEA)-neurochips. We have extensive experience with a range of primary neuronal cell cultures and human neuronal stem cells, and with the recording and mathematical analysis of their spontaneous and compound-induced network activities.

Using our proprietary pattern recognition algorithms, we determine the similarity or difference of a compound to other known compounds.

This allows screening for differentiation from marketed compounds, mimicry of endogenous ligands, and assessment of synergistic effects. The cultures also allow to assess effects on neuronal maturation, synaptogenesis, and regenerative properties as well as assessing the differentiation process of human stem cells cultures.

Selected Publications:

  • Hammer, H. et al. A multifaceted GABAA receptor modulator: Functional properties and mechanism of action of the sedative-hypnotic and recreational drug methaqualone (Quaalude®). Molecular Pharmacology, 2015 Jun 8. pii: mol.115.099291.
  • Schulz L. et al. Evaluation of drug-induced neurotoxicity based on metabolomics, proteomics and electrical activity measurements in complementary CNS in vitro models. Toxicol In Vitro. 2015 Dec 25;30(1 Pt A):138-65. doi: 10.1016/j.tiv.2015.05.016. Epub 2015 May 27.
  • Katsumata S. et al. Electrophysiological Characterization of GABA-A alpha5 negative allosteric modulators in hippocampal neurons. Proceedings: 31st International Conference of Alzheimer's Disease International (ADI) 21-24 April 2016, Budapast, Abstract book, p.230, [http://www.adi2016.org/images/downloads/adi2016_abstract_book.pdf]
  • Gramowski-Voß A. et al. Enhancement of cortical network activity in vitro and promotion of GABAergic neurogenesis by stimulation with an electromagnetic field with a 150 MHz carrier wave pulsed with an alternating 10 and 16 Hz modulation. Front. Neurol. 2015. 6:158. doi: 10.3389/fneur.2015.00158
  • Parenti C. et al. The multitarget opioid ligand LP1's effects in persistent pain and in primary cell neuronal cultures. Neuropharmacology 2013 Aug;71:70-82.
  • Johnstone AFM. et al. Microelectrode arrays: A physiologically based neurotoxicity testing platform for the 21st century. Neurotoxicology 2010 Aug;31(4):331-50.
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Cell Electrophysiology
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Ex vivo Electrophysiology
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Cytotoxicity Assays
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In combination with our MEA analysis or as stand-alone service we provide a complete report for toxicity profiling and safety screening including information about acute or chronic cell viability and cytotoxicity assessed by:

  • MTT assay (enpoint analysis)
  • LDH assay (non-invasive multiwell analysis of culture medium)
    -... Show more »

In combination with our MEA analysis or as stand-alone service we provide a complete report for toxicity profiling and safety screening including information about acute or chronic cell viability and cytotoxicity assessed by:

  • MTT assay (enpoint analysis)
  • LDH assay (non-invasive multiwell analysis of culture medium)
  • Necrosis assay (CytoTox™ DNA staining)
  • Apoptosis assay (Annexin-V)
  • Proliferation assay (EdU staining)
  • Extracellular protease detection assay (non-invasive multiwell analysis of culture medium)
  • Customized cell stainings:
    • specific loss of neurons, glia cells or sub-populations (e.g. GABAergic, dopaminergic neurons),
    • neurite detection and quantification
    • synapse numbers
    • activation of microglia
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Cell Viability Assays
Price on request

In combination with our MEA analysis or as stand-alone service we provide a complete report for toxicity profiling and safety screening including information about acute or chronic cell viability and cytotoxicity assessed by:

  • MTT assay (enpoint analysis)
  • LDH assay (non-invasive multiwell analysis of culture medium)
    -... Show more »

In combination with our MEA analysis or as stand-alone service we provide a complete report for toxicity profiling and safety screening including information about acute or chronic cell viability and cytotoxicity assessed by:

  • MTT assay (enpoint analysis)
  • LDH assay (non-invasive multiwell analysis of culture medium)
  • Necrosis assay (CytoTox™ DNA staining)
  • Apoptosis assay (Annexin-V)
  • Proliferation assay (EdU staining)
  • Extracellular protease detection assay (non-invasive multiwell analysis of culture medium)
  • Customized cell stainings:
    • specific loss of neurons, glia cells or sub-populations (e.g. GABAergic, dopaminergic neurons),
    • neurite detection and quantification
    • synapse numbers
    • activation of microglia
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Drug Target Validation
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Biochemical validation

  • Western blotting
  • Customized cell staining

Please note: we also prepare cell lysates or fixed cell cultures for you for internal target validation

Electrophysiological validation

In our blocker experiments we block your target-of-interest with a specific blocker (either purchased or... Show more »

Biochemical validation

  • Western blotting
  • Customized cell staining

Please note: we also prepare cell lysates or fixed cell cultures for you for internal target validation

Electrophysiological validation

In our blocker experiments we block your target-of-interest with a specific blocker (either purchased or blinded) by directly combining with your compounds in a combination experiments.

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Neurogenesis
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MEA neurochips are a valuable tool to detect compound effects on neuronal maturation. By use of the MEA-neurochip technology we can directly monitor and characterize maturation of primary mouse or human iPSC-derived network activity in electrically active neuronal networks.

Compound application during maturation (either single... Show more »

MEA neurochips are a valuable tool to detect compound effects on neuronal maturation. By use of the MEA-neurochip technology we can directly monitor and characterize maturation of primary mouse or human iPSC-derived network activity in electrically active neuronal networks.

Compound application during maturation (either single or repeated) can affect the onset of spontaneous network activity and the maturation of both individual neurons and whole network characteristics.

We assay the electrical activity patterns of dissociated primary neurons in culture from an early embryonic stage up to 6 weeks of maturation in vitro. The activity fingerprint is described by more than 200 parameters, which allows a sensitive measure for functional maturation and the effects of biologics and chemical compounds thereon to assess effects on synaptogenesis and neurogenesis.

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Drug Combination Activity Testing
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The combination of new compounds with marketed drugs for combination therapy is a promising approach to generate new therapeutic opportunities. The identification of the ideal combination of compounds at an optimal ratio and concentration requires a large number of sensitive tests.

NeuroProof offers a cost-effective and elegant... Show more »

The combination of new compounds with marketed drugs for combination therapy is a promising approach to generate new therapeutic opportunities. The identification of the ideal combination of compounds at an optimal ratio and concentration requires a large number of sensitive tests.

NeuroProof offers a cost-effective and elegant assay to screen compound combinations with a phenotypic read out.

Using NeuroProof's complex data analysis it is possible to detect new and unexpected effects in drug combinations which can be exploited for new therapeutic applications. The precise and highly reproducible data allows the exact determination of optimal ratios of compound mixtures.

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Stem Cell Assays
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Human neuronal stem cell and iPS cell cultures are of high scientific and practical interest in drug discovery, toxicology and for future therapeutic applications. To understand the differentiation process of these stem cell cultures into neurons is an important and complex task and an essential prerequisite for the functional... Show more »

Human neuronal stem cell and iPS cell cultures are of high scientific and practical interest in drug discovery, toxicology and for future therapeutic applications. To understand the differentiation process of these stem cell cultures into neurons is an important and complex task and an essential prerequisite for the functional characterization of test compounds in a human cellular background.

Our offer in service and FTE projects

  • Phenotypic screening of test compounds on human iPSC derived neurons: We have developed and established protocols for human iPSC neurons on MEAs (partly together with our commercial partners who offer iPSC-neurons and NPCs).
  • Functional characterization of your hiPSC-derived neurons or progenitors: We apply your protocol for culturing neurons or NPCs and screen them during their synaptic and neurogenic development on micro-electrode arrays. Having more than four years of experience with hiPSC neuron recordings, we optimize your culture protocols based on functional activity readout and compare activity with commercially available reference cell lines.
  • Comparing functional human and mouse phenotype: We have a database of several hundreds to thousands of datasets from mouse tissue-specific neuron cultures. We compare stem cell-derived neuronal network activity patterns with primary cell recordings. This is a powerful tool to characterize tissue-, species- and compound-specific functionality.
  • Parkinson’s disease in a dish: We induce disease-associated functional phenotypes into human iPSC neuronal networks for the screening of neuro-protective compounds. We use dopaminergic neuronal networks and functionally impaired them e.g. by recombinant monomeric or filamentous alpha synuclein or low-concentrated MPP+. These disease phenotypes are quantified by multi-parametric data analysis and sophisticated artificial intelligence tools. This allows us to rank rescue efficacy of your compounds in a simple manner.
  • Functional biomarker screening: we compare human iPSC-derived neurons from healthy individuals with patient-derived cultures and identify disease-related functional parameters. These functional biomarkers allow specific compound screening.
  • Together with you we establishment specific disease models based on your specific indication and gene of interests.
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Microelectrode Array (MEA) Data Analysis
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NeuroProof offers its capabilities as a service to analyze customer's electrophysiological data from microelectrode array setups. Customers can benefit from our experience which we gained during the last 15 years. The service includes in vivo and in vitro studies with microelectrode array setups, quality analyses of spike... Show more »

NeuroProof offers its capabilities as a service to analyze customer's electrophysiological data from microelectrode array setups. Customers can benefit from our experience which we gained during the last 15 years. The service includes in vivo and in vitro studies with microelectrode array setups, quality analyses of spike train data and action potential wave form analyses.

After a first inspection of the customer's data we can propose a data analysis concept which fits customer’s requests. If needed, spike train data can be reanalyzed by off line unit separation. Spike train data can be characterized by over 200 activity describing parameters, covering changes in the general activity, the burst structure, the oscillatory behavior, network synchronicity and connectivity. By statistical analysis on the basis of multivariate statistics customers gain reproducible and valid results, supporting their drug discovery and drug development process.

Additionally, NeuroProof offers the development of customized software applications for MEA derived data.

Our services cover all levels of MEA-neurochip data analysis; spike train analysis, multivariate data analysis and building up of in house data bases.

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In vitro Disease Models
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Our contribution to successful drug development for the treatment of neuro degenerative diseases is the offer of relevant in vitro disease models which are based on a functional and phenotypic neuronal readout.

These models can be used for hit identification and validation, lead selection and lead optimization. Due to the... Show more »

Our contribution to successful drug development for the treatment of neuro degenerative diseases is the offer of relevant in vitro disease models which are based on a functional and phenotypic neuronal readout.

These models can be used for hit identification and validation, lead selection and lead optimization. Due to the functional phenotypic readout, compounds can be investigated independently of their main and potentially unknown target.

We offer the following functional in vitro assays:

Alzheimer:

Functional Abeta42 protection assay: acute treatment of primary hippocampus neurons with Abeta42.

  • Acute phenotypic rescue efficacy of Abeta challenged neurons.
  • Time-dependent phenotypic rescue efficacy of Abeta-challenged neurons to elucidate the temporal stability of rescue.
  • Acute phenotypic prevention efficacy before challenging with Abeta.

Parkinson:

Functional MPP+ protection assay: challenge of developing primary midbrain dopaminergic neurons by single, low concentration MPP+

  • phenotypic rescue efficacy of MPP+ challenged neurons.
  • phenotypic prevention efficacy before challenging with MPP+.

We offer these models for the evaluation of your compounds on a fee-for-service basis, providing excellent reproducibility and highly significant results for established reference compounds.

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In vitro Epilepsy Models
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Epilepsy is one of the most common neurological disorders worldwide. The term epilepsy comprises a variety of different syndromes which all have the occurrence of recurrent unprovoked seizures in common.

Although most patients become seizure-free on antiepileptic drugs, about 20-30% of patients continue to have seizures. For... Show more »

Epilepsy is one of the most common neurological disorders worldwide. The term epilepsy comprises a variety of different syndromes which all have the occurrence of recurrent unprovoked seizures in common.

Although most patients become seizure-free on antiepileptic drugs, about 20-30% of patients continue to have seizures. For those patients no adequate pharmacological treatment is available yet.

Current treatment of epilepsy with drugs relies on symptomatic control of seizures. However, epilepsy cannot be cured with medication, because the mechanisms underlying epileptogenesis are still largely unknown.

NeuroProof has developed an in vitro screening assay to investigate the effects of new classes of antiepileptic compounds. Neuronal networks of embryonic hippocampus neurons grow on MEA chips and develop spontaneous ictal-like activity under certain culture conditions. This activity is characterized by synchronous burst events with a duration of >5 s. Anticonvulsive compounds such as carbamazepine and valproic acid have been shown to resolve these long burst events, while duration of bursts from healthy hippocampus networks are not affected by these compounds.

The NeuroProof technology can also help to understand pro-convulsive compounds such as picrotoxin that induce ictal-like discharges in network activity of frontal cortex cultures. During these ictal discharges bursts are arranged into longer duration burst events that intermit frontal cortex activity.

Using our classification analysis we can predict excitatory or only pro-convulsive potential of test compounds.

NeuroProof offers specific in vitro assays for drug discovery in epilepsy. Human iPSC derived neurons can also be used in these assays to increase translation into humans.

The NeuroProof technology can also help to understand pro-convulsive compounds such as picrotoxin that induce ictal-like discharges in network activity of frontal cortex cultures. During these ictal discharges bursts are arranged into longer duration burst events that intermit frontal cortex activity.

Using our classification analysis we can predict excitatory or only pro-convulsive potential of test compounds. Here we refer to our side effect assays.

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Microelectrode Array (MEA) Recordings
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We perform phenotypic screening service to assess a compound's action in a micro-circuit including a multitude of physiological relevant receptors with primary neurons or human stem cell-derived cultures growing on a micro electrode array chip. We compare the functional fingerprint of your new compound with profiles of your... Show more »

We perform phenotypic screening service to assess a compound's action in a micro-circuit including a multitude of physiological relevant receptors with primary neurons or human stem cell-derived cultures growing on a micro electrode array chip. We compare the functional fingerprint of your new compound with profiles of your reference compounds or those in our substance database.

This allows:

  • Identifying the neuroactive concentration ranges based on multi-parametric concentration-response data (picomolar to micromolar)
  • Differentiating your new compounds against target-specific substances, endogenous ligands and standard of care drugs.
  • Gaining insight into the functional similarity to reference compounds at certain concentrations (e.g. therapeutic concentration of more than 30 clinical CNS drugs already tested by NeuroProof). This analysis is performed in a concentration-specific manner for the test compound.
  • Proposing clinical indication by sorting the profile into several drug classes (e.g. anticonvulsants, antipsychotics, antidepressants, pro-cognitives, analgesics and more).
  • Assess potential repurposing opportunities for your compounds which might rescue your compounds which development was stopped due to safety or efficacy concerns.
  • Elucidate potential side effects such as pro-convulsive or sedative (read: prediction of side effect).

We deliver:

  • The phenotypic profiling which characterizes a compound's action in the whole concert of all receptors it affects. Our experience shows that each compound exhibits a distinct and quantitatively assessable profile with unique properties.
  • A characterization of a compound's action established by our comprehensive multivariate data analysis. You receive a description of your compound's action on neuronal networks with a multitude of parameters (general activity, regularity, burst structure/strength, synchronicity/connectivity/communication).
  • An overall comparison to your reference compounds or known compounds of our database.

The following primary cell cultures are available:

  • frontal cortex
  • hippocampus
  • amygdala/hippocampus co-culture
  • midbrain
  • midbrain/cortex co-culture
  • spinal cord
  • hypothalamus
  • human induced pluripotent stem cell-derived neurons
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Cell Viability & Proliferation Assays
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Toxicology
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Drug Discovery
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Drug Discovery & Development
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Cells and Tissues
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Biology
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Pharmacology & Toxicology
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Lead Identification and Validation
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In vitro Toxicity Testing
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Functional & Cell Type Specific Assays
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Cell-Based Assays
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Neuroscience
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