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Open Access Green as soon as Postprint is submitted to ZB.
Adapting human pluripotent stem cells to high-throughput and high-content screening.
Nat. Protoc. 8, 111-130 (2013)
The increasing use of human pluripotent stem cells (hPSCs) as a source of cells for drug discovery, cytotoxicity assessment and disease modeling requires their adaptation to large-scale culture conditions and screening formats. Here, we describe a simple and robust protocol for the adaptation of human embryonic stem cells (hESCs) to high-throughput screening (HTS). This protocol can also be adapted to human induced pluripotent stem cells (hiPSCs) and high-content screening (HCS). We also describe a 7-d assay to identify compounds with an effect on hESC self-renewal and differentiation. This assay can be adapted to a variety of applications. The procedure involves the culture expansion of hESCs, their adaptation to 384-well plates, the addition of small molecules or other factors, and finally data acquisition and processing. In this protocol, the optimal number of hESCs plated in 384-well plates has been adapted to HTS/HCS assays of 7 d.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Patient-specific Ipscs ; Human Somatic-cells ; Term Self-renewal ; Small Molecules ; Suspension-culture ; In-vitro ; Human Blastocysts ; Dopamine Neurons ; Scalable Culture ; Defined Factors
ISSN (print) / ISBN
1754-2189
e-ISSN
1750-2799
Journal
Nature Protocols
Quellenangaben
Volume: 8,
Issue: 1,
Pages: 111-130
Publisher
Nature Publishing Group
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Developmental Genetics (IDG)