Neurotoxicity Screening and Testing

The majority of studies performed by HemoGenix® incorporate a Complete Service, Full Report that can be used for an IND application. More recently, Sponsors have requested a more streamlined study and studies involving high-throughput screening. For this reason, HemoGenix® now provides 3 types of study format. 

1. Complete Service, Full Report Study: Fully customized study that includes the Study Plan, Draft Text and Final Text Report with QA audit.
2. Rapid Toxicity Study: A customized study that includes the Study Plan and full protocol, raw results and graphical data in a single Excel Workbook. No formal text report and no QA audit is performed. This type of study is designed for early drug development. No interpretation or conclusions are provided.
3. High-Throughput Screening Module (presently only available for human studies): A screening study is designed for high-throughput screening of compounds during ADME/Tox screening in multiples of 5 on specific cell populations to provide the most important ranking information. The full protocol, raw results and graphical data are provided in an Excel Workbook. No interpretation or conclusions are provided. This type of Screening Study is part of the ComparaTOX™ HT Platform.

There are many different types of neural/nuronal cells available, but not all are suited for cytotoxicity studies. The primary source of cells used for cytotoxicity studies, including high-throughput neurotoxicity screening studies are:

• iPS-derived human iCell GABANeurons from Cellular Dynamics
• iPS-derived human iCell Motor Neurons from Cellular Dynamics. 

For other cell types and species used for in vitro neurotoxicity testing, please contact HemoGenix^®.

Most in vitro neurotoxicity studies are performed on human iPS-derived cells in pre-coated 96- or 384-well plates. The assay platform used is called NeuroGlo™-Tox HT and is a proprietary assay based originally on the HemoGenix^® HALO^® Platform for stem and progenitor cell hematotoxicity screening and testing. Like the HALO® Platform, NeuroGlo™-Tox HT incorporates the most sensitive non-radioactive signal detection system available, namely ATP bioluminescence. 

Using NeuroGlo™-Tox HT for in vitro neurotoxicity screening (see below) and testing, has the following benefits:

• Incorporates the most sensitive ATP bioluminescence readout available to measure cytotoxicity.
• NeuroGlo™-Tox HT is a calibrated and fully standardized assay platform.
• The NeuroGlo™-Tox HT readout has been validated according to FDA Bioanalytical Method Validation Guidelines and incorporates measurement assurance parameters to ensure results are trustworthy.
• NeuroGlo™-Tox HT is available in 96- or 384-well plate formats.
• The standardized cytotoxicity assay involves a 48-72hr drug treatment protocol.
• Designed for multiplexing with other assay using the same sample (see below).

NeuroGlo™-Tox HT has been developed for high-throughput screening typically used during primary and secondary ADME/Tox drug development.  

Besides normal high-throughput screening with a Full Report, HemoGenix® also provides a Rapid High-Throughput Screening Module for multiples of 5 compounds using iCell GABANeurons as the target cells. These studies are meant to provide raw data in the shortest period in an Excel Workbook.

Similar rapidly produced results can also be obtained for comparing different tissues for multiples of 5 compounds using the HemoGenix^® ComparaTOX™ HT Platform that allows compounds to be ranked in order of toxicity. See more information on ComparaTOX™. 

NeuroGlo™-Tox HT can be multiplexed with a variety of other assays or add-on assays can be used for mechanism of action (MOA) studies. These might include, but are not limited to:

• NeuroGlo™ Real Time for growth kinetic analysis and time of onset of cytotoxicity.
• Resazurin/resorufin, non-lytic fluorescence assays.
• Tetrazolium WST, non-lytic absorbance assays.
• Membrane integrity: LDH and/or PI
• Biochemical apoptosis assays: Caspases
• Flow cytometry of membrane markers that correlate with cell function.
• OxyFLOW: Oxidative DNA Damage
• Oxidative stress: GSH
• Mitochondrial dysfunction: Mitochondrial ToxGlo™

For in-house in vitro neurotoxicity testing or functional studies on neural cells, assay kits from Preferred Cell Systems™ are available. 

NeuroGlo™-Tox HT: For in-house neurotoxicity testing. Please contact Preferred Cell Systems™ for Complete Assays that include cryopreserved cells.

NeuroGlo™: ATP bioluminescence functional assays for neural cells.

NeuroFluor™: Resazuring/Resorufin fluorescence, non-lytic, functional assays for neural cells.

NeuroLight™: tetrazolium WST absorbance non-lytic, functional assays for neural cells.