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

• Published in: Journal of biomolecular screening Vol. 18; no. 3; p. 258
• Main Authors: McLaren, Donna, Gorba, Thorsten, Marguerie de Rotrou, Anita, Pillai, Gopalan, Chappell, Clare, Stacey, Alison, Lingard, Sarah, Falk, Anna, Smith, Austin, Koch, Philipp, Brüstle, Oliver, Vickers, Richard, Tinsley, Jon, Flanders, David, Bello, Paul, Craig, Stewart
• Format: Journal Article
• Language: English
• Published: United States 01-03-2013
• Subjects: Cell Culture Techniques - methods; Cell Proliferation - drug effects; Cell Survival - drug effects; Cells, Cultured; Culture Media, Serum-Free; Cyclin-Dependent Kinase 2 - metabolism; Dose-Response Relationship, Drug; Drug Evaluation, Preclinical - methods; Follow-Up Studies; High-Throughput Screening Assays - methods; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Neural Stem Cells - cytology; Neural Stem Cells - drug effects
• ISSN: 1552-454X
• DOI: 10.1177/1087057112461446

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.