Assay to test compound-derived impairment in neurite outgrowth in human iPSC-derived immature dorsal root ganglia (iDRG) neurons (PeriTox; UKN5)

This in vitro test method is based on human iPSC-derived immature dorsal root ganglia (iDRG) neurons at a stage of neurite growth. It assesses (a) disturbances in the development of the (peripheral) nervous system, and (b) direct damage to the peripheral nervous system, by exposure to toxicants. The neurite area (which serves as indirect measurement of neuronal interconnectivity) of stained differentiating neurons, as well as cellular viability are measured simultaneously using high content imaging. The processes of neurite outgrowth and cell death are measured. The data of this method are meant to predict (a) developmental disorders in children caused by compound exposure during fetal development, and (b) damage to the developed nervous system, in particular to the peripheral nervous system. The method predicts neurotoxicity. It has been used in the screening of medium-sized compound libraries, and has undergone some mechanistic evaluation, and been linked to AOP-279 (AOPwiki ID) / ETR09N (EU-ToxRisk AOP task ID) (Peripheral neuropathy caused by microtubule interacting drugs).

Descriptions of Assays

Our assays cover crucial functional endpoints for (developmental) neurotoxicity testing:

• Assay to test viability and impairment of migration of human neural crest cells (cMINC; UKN2);
• Assay to test compound-derived impairment in neurite outgrowth in human mature dopaminergic neurons (NeuriTox; UKN4);
• Assay to test compound-derived impairment in neurite outgrowth in human iPSC-derived immature dorsal root ganglia (iDRG) neurons (PeriTox; UKN5).
• Neurofunctional assays based on receptor interactions, for dopamine transporter and assessing network formation (e.g. assessment of Ca²+ signaling and electrical activity on multi-electrodes arrays);
• Assays for neuroprotective drug candidates in conventional cell cultures and complex organoids.

Services provided include:

• Use of in vitro tests for the assessing of developmental toxicity (DT) and developmental neurotoxicity (DNT) hazard.
• Use of in vitro tests for the assessing of neurotoxicity (NT) hazard
• Use of in vitro tests for assessing efficacy and mode of action (MoA) of neuroprotective drugs.
• Complementation of in vivo studies for NT, DNT, DT to provide important information on MoA of chemicals.

Vendor: CAAT-Europe