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
(2013) In Journal of Biomolecular Screening 18(3). p.68-258- Abstract
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... (More)
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.
(Less)
- author
- publishing date
- 2013-03
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- 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, Neural Stem Cells/cytology
- in
- Journal of Biomolecular Screening
- volume
- 18
- issue
- 3
- pages
- 11 pages
- publisher
- SAGE Publications
- external identifiers
-
- pmid:23042076
- scopus:84873669503
- ISSN
- 1087-0571
- DOI
- 10.1177/1087057112461446
- language
- English
- LU publication?
- no
- id
- 861b1f58-87e7-4b66-9155-3529b9dc49ec
- date added to LUP
- 2021-08-10 13:33:35
- date last changed
- 2024-04-06 07:45:56
@article{861b1f58-87e7-4b66-9155-3529b9dc49ec,
abstract = {{<p>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.</p>}},
author = {{McLaren, Donna and Gorba, Thorsten and Marguerie de Rotrou, Anita and Pillai, Gopalan and Chappell, Clare and Stacey, Alison and Lingard, Sarah and Falk, Anna and Smith, Austin and Koch, Philipp and Brüstle, Oliver and Vickers, Richard and Tinsley, Jon and Flanders, David and Bello, Paul and Craig, Stewart}},
issn = {{1087-0571}},
keywords = {{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; Neural Stem Cells/cytology}},
language = {{eng}},
number = {{3}},
pages = {{68--258}},
publisher = {{SAGE Publications}},
series = {{Journal of Biomolecular Screening}},
title = {{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}},
url = {{https://lup.lub.lu.se/search/files/101076858/Automated_Large_Scale_Culture_.pdf}},
doi = {{10.1177/1087057112461446}},
volume = {{18}},
year = {{2013}},
}