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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.
Impact Factor
Scopus SNIP
Web of Science
Times Cited
Times Cited
Scopus
Cited By
Cited By
Altmetric
7.960
3.242
50
54
Anmerkungen
Besondere Publikation
Auf Hompepage verbergern
Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
Patient-specific Ipscs ; Human Somatic-cells ; Term Self-renewal ; Small Molecules ; Suspension-culture ; In-vitro ; Human Blastocysts ; Dopamine Neurons ; Scalable Culture ; Defined Factors
Sprache
englisch
Veröffentlichungsjahr
2013
HGF-Berichtsjahr
2013
ISSN (print) / ISBN
1754-2189
e-ISSN
1750-2799
Zeitschrift
Nature Protocols
Quellenangaben
Band: 8,
Heft: 1,
Seiten: 111-130
Verlag
Nature Publishing Group
Begutachtungsstatus
Peer reviewed
Institut(e)
Institute of Developmental Genetics (IDG)
POF Topic(s)
30204 - Cell Programming and Repair
30204 - Cell Programming and Repair
Forschungsfeld(er)
Genetics and Epidemiology
Genetics and Epidemiology
PSP-Element(e)
G-500500-003
G-500500-001
G-500591-001
G-500500-001
G-500591-001
PubMed ID
23257981
WOS ID
WOS:000313051300009
Scopus ID
84872013374
Erfassungsdatum
2013-02-01