Sponsored by Alpha MED Scientific and Axol Bioscience
"Exploring the latest advancements in neural stem cell technology and the most sensitive high- throughput multi-electrode array platform, leading to a better understanding of neuronal networks"
Monday November 13, 2017
Renaissance Washington, DC Downtown Hotel, Congressional Ballroom A
Neurons in vitro can provide valuable insights into the mechanisms of the behavior and disease. The intricacies of in vitro assays require a high-fidelity system to fully characterize activity. The MED64 is a high-fidelity microelectrode array (MEA) that is engineered to detect a broad range of signals. In this symposium, researchers will demonstrate the power of high sensitivity MEAs for measuring activity in stem cell derived neurons and evoked field potentials in multiple acute slices.
Alfredo Cabrera-Socorro, PhD
Janssen Research & Development, Johnson & Johnson
|Towards the standardization of iPSC technology for drug development in neuroscience: challenging data reproducibility using iPSC-derived cells ( 1 )|
Ryan Arant, PhD
|High-throughput Electrophysiology for Brain Slice Plasticity - how to quadruple your output ( 2 )|
Ass. Prof Ikuro Suzuki, PhD
Tohoku Institute of Technology
|Evaluation of drug-induced neurotoxicity in human iPSC-derived cortical and sensory neurons using high-throughput MEA system ( 3 )|
Prof. Glen Kisby, PhD
Western University of Health Sciences
|Cellular models of human neurodevelopment differ in their sensitivity to environmental chemicals ( 4 )|
Run by the University of Saskatchewan
Kinome Profiling of Neural Stem cells (NSC) derived from (induce pluripotent stem cells (iPSC) of Huntington's disease patient
Session Date and Time
: Sunday, November 12, 1:00 PM - 5:00 PM
Presenter at Poster : 3:00 PM - 4:00 PM
Location : Halls A-C
Number : 212.03 / 03
Authors : A. Baharani, E. Scruten, S. Napper
The researchers utilized Huntington's Disease (HD) iPSC-derived human neural progenitor cells differentiated in Axol's innovation lab to identify key signal transduction proteins that are dysregulated in HD via Kinome analyses.
1. The induced pluripotent stem cells (iPSCs) technology is not yet a fully consolidated pre-clinical model within pharmaceutical drug discovery programs. Duration of the differentiation process and the challenge to generate fully mature neurons has delayed application for drug development in neuroscience. The translational value of iPSC technology can only be demonstrated by developing models which reproducibility is demonstrated across laboratories. Here we will discuss current efforts between academic and industry partners across Europe aiming to establish reliable iPSC-based models of neurodegeneration.
3. The functional network of human induced pluripotent stem cell (hiPSC)-derived neurons is a potentially powerful in vitro model for evaluating drug toxicity. Epileptiform activity is one of toxicological phenomena in central nervous system. We evaluated the dynamics of epileptiform activities and the effect of anti-convulsant drug in cultured hiPSC-derived cortical neurons (Axol Bioscience) using high-throughput multielectrode array (MEA) system. Electrophysiological seizes were induced by 4-Aminopyridine, Pentylentetrazole, Pilocarpine, and Chlorpromazine in a concentration-dependent manner. Anti-convulsant phenytoin suppressed induced epileptiform activity. We analyzed the MEA data using multi-parameters and separated responses depending on the drug types. In addition, we detected the pain responses in cultured hiPSC-derived sensory neurons. Firstly, we confirmed hiPSC-derived sensory neurons expressed typical sensory neural marker Nav1.7, TRPV1, and TRPA1 using immunostaining. Next, we detected the physiological responses to temperature change, capsaicin, menthol, and wasabi by change of spike rate using MEA system. Human iPSC-derived neurons were classified into 27 types depending on physiological responses against 3 compounds. We also detected the responses to anti-cancer drugs oxaliplation and vincristine. Our results suggested that electrophysiological measurement in cultured hiPSC derived sensory neurons using MEA system are suitable to toxicological assay for epileptiform activities in CNS and pain responses in PNS.
4. Early life exposure to environmental chemicals is an important risk factor for neurodevelopmental disorders. Human neuroblastoma cell lines (e.g., SY5Y) are the most widely used in vitro model for evaluating the neurotoxic properties of environmental chemicals while human neuroprogenitor cells (hNPCs) are a newer and less used model. We compared the sensitivity of both models at different developmental stages (i.e., immature vs. mature) to the anti-seizure drug valproic acid (VPA), the plastic and resin contaminant bisphenol-A (BPA), and the carcinogen methylazoxymethanol (MAM). Immature SY5Y cells and hNPCs were treated with various concentrations of VPA, BPA, or MAM for 24 hr and then evaluated for cell viability (Calcein-AM) and mitochondrial function (PrestoBlue®). A similar set of treated SY5Y cells and hNPC cultures were differentiated with various growth factors for 10-24 days (mature cells) before measuring cell viability and mitochondrial function. Significant differences were observed in the sensitivity of SY5Y cells and hNPCs to the three neurotoxins. VPA was more toxic to mature hNPCs than comparably treated immature or mature SY5Y cells (p<0.05), but the effect was independent of concentration. Like VPA, BPA was more toxic to mature hNPCs than comparably treated immature or mature SY5Y cells (p<0.05), but the effect was concentration dependent (p<0.05). MAM was also more toxic to mature hNPCs than comparably treated immature hNPCs or mature SY5Y cells (p<0.05), but like BPA, the effect was concentration dependent. Thus, these in vitro human neural models show significant developmental and concentration-dependent differences after treatment with environmental chemicals. These differences might be due to the disruption of distinct mechanisms in each neurodevelopmental model. These findings raise important questions about the use of established neural culture models (i.e., SY5Y) to predict developmental neurotoxicity of environmental chemicals in humans.