期刊
JOURNAL OF BIOMOLECULAR SCREENING
卷 18, 期 3, 页码 258-268出版社
SAGE PUBLICATIONS INC
DOI: 10.1177/1087057112461446
关键词
neural stem cells; automation; assay development; proliferation; screening
资金
- European Commission [LSHB-CT-2007/037766, HEALTH-F5-2010-266753]
- state of North Rhine Westphalia (BIO.NRW)
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.
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