Automated large-scale culture and medium-throughput chemical screen for modulators of proliferation and viability of human induced pluripotent stem cell-derived neuroepithelial-like stem cells.

The aim of this study was to demonstrate proof-of-concept feasibility for the use of human neural stem cells (NSCs) for high-throughput screening (HTS) applications. For this study, an adherent human induced pluripotent stem (iPS) cell-derived long-term, self-renewing, neuroepithelial-like stem (lt-NES) cell line was selected as a representative NSC. Here, we describe the automated large-scale serum-free culture ("scale-up") of human lt-NES cells on the CompacT SelecT cell culture robotic platform, followed by their subsequent automated "scale-out" into a microwell plate format. We also report a medium-throughput screen of 1000 compounds to identify modulators of neural stem cell proliferation and/or survival. The screen was performed on two independent occasions using a cell viability assay with end-point reading resulting in the identification of 24 potential hit compounds, 5 of which were found to increase the proliferation and/or survival of human lt-NES on both occasions. Follow-up studies confirmed a dose-dependent effect of one of the hit compounds, which was a Cdk-2 modulator. This approach could be further developed as part of a strategy to screen compounds to either improve the procedures for the in vitro expansion of neural stem cells or to potentially modulate endogenous neural stem cell behavior in the diseased nervous system.

A rapid, high content, in vivo model of glucocorticoid-induced osteoporosis.

Glucocorticoid-induced osteoporosis (GIOP) is a major clinical problem given the widespread use of steroids and limited efficacy of biphosphonates. Existing animal models of GIOP are both slow and expensive. Hence, there is a need both for adjunctive modelling systems, as well as more efficacious therapies for the treatment of GIOP. We have addressed this issue through the creation of a zebrafish model of GIOP, which can be used for 96-well plate in vivo screening with an assay time of 5 days. The model demonstrates key similarities to human GIOP including a partial response to bisphosphonates. The ability to extract detailed pharmacological data, including concentration-response analyses, enables the screening and ranking of candidate therapeutic compounds. In addition, the zebrafish model is highly relevant for pathway dissection through genetic knockdown and overexpression studies.