Ambrose, Jessi, Mapa, Felipa, Ross, Nathan, Chen, Julie, Ye, Chaoyang and Bilican, Bilada (2017) Automated Scale-up and Maintenance of Stem Cell-Induced Neurons for Long-Term Culture and High-Throughput Screening. SLAS Technology.

Abstract

Successful drug screens benefit from testing large compound libraries in biologically relevant models. Recent advancements in the reprogramming of somatic cells to pluripotent stem (iPS) cells and the methods to differentiate such cells into tissue-specific cell types have allowed for increased reliability and scale-up of more relevant cell models. Specifically, the method of inducing iPSC and ESCs to cortical excitatory neurons by expression of NGN2 has made scale-up for HTS possible, but challenges such as increased risk of contamination, stability of growth media, plate coatings, and frequent media changes remain. Here we present a method for expanding and differentiating iPSC and ESCs into induced neurons (iNs) using a CompacT SelecT for up to 48 days and the subsequent plating and maintaining of the cells in 1536 well plates using a GNF Systems automated screening system for up to 14 days. We were able to demonstrate that cells cultured using these methods maintained the phenotype of NGN2-induced neurons through expression profiling with single-cell qPCR and protein expression and localization by immuno-fluorescence. We were further able to show that iN cells frozen after a 48 hour induction can be re-plated into 1536 well plates and maintain the same gene expression profiles, allowing for batched cell banking. These methods allow for the successful scale-up and long-term culture of iN cells to enable HTS.

Item Type:	Article
Date Deposited:	17 Feb 2017 00:45
Last Modified:	17 Feb 2017 00:45
URI:	https://oak.novartis.com/id/eprint/31725