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The use of FCDI iCell neurons and supplements is essential for 2D neuronal cultures used in neurotoxicity screening assays for siRNA or ASO toxicity projects. For some 2.5 years, NCATS scientists have used this cell line, provided by Fujifilm Cellular Dynamics, to optimize both 2D MEA motor neuron culture conditions (POTS: 22-004194) that generate consistent and reproducible baseline measurements and have established a system. The iPSC technology has only existed for some 10 years, and this established system is extremely sensitive.
A change in the source of these neurons would be catastrophic. It would significantly alter such parameters as growth rates, onset and levels of spontaneous activity, potentially requiring critical last-minute changes, necessitating additional testing, optimization of experimental protocols, and costs that will far outweigh the cost of this requirement. Use of FCDI iCells also allows scientists to use ihPSCs that are derived from the same patient from cells used in previous experiments, lowering the chance that genetic diversity will cause variability in assays using these cells.
Previous optimization of both neuronal 2D (POTS: 24-002733) cultures have established the optimal number of neurons for both types of cultures to screen ASOs for neurotoxicity and have been used to successful complete early Milestone of the ongoing this project In order to complete the HEAL collaborative project hiPSC these specific neurons and supplements for 2D culture plates are needed and is essential for the continuity of science.
The Therapeutic Development Branch (TBD) within the National Center for Advancing Translational Sciences (NCATS) Division of Preclinical Innovation (DPI) collaborative project Under HEAL Initiative is aimed at developing assays for predictive toxicity of siRNA or antisense oligonucleotides (ASOs). 2D cultures using human induced pluripotent stem cell (hiPSC)-derived neurons (from non-fetal tissue source) have been adapted and extensively optimized by NCATS scientist to successfully predict neurotoxicity of siRNA or ASOs.
2D ihPSC-derived spinal motor neurons co-cultured with astrocytes on 48-well microelectrode array (MEA) plates allows spontaneous electrical activity to be measured over the course of the culture. Changes in neuronal firing following siRNA/ASO treatment can be quantified to determine neurotoxicity and is typically used as a secondary screen for hits detected with 3D cultures.
Previous optimization of both neuronal 2D (POTS: 24-002733) cultures have established the optimal number of neurons for both types of cultures to screen ASOs for neurotoxicity. In order to complete the HEAL project, enough hiPSC neurons and supplements for 2D culture plates are needed and is a requirement to meet the goals of this project.
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