High-throughput screening assay for the identification of compounds regulating self-renewal and differentiation in human embryonic stem cells.
High-throughput screening (HTS) of chemical libraries has become a critical tool in basic biology and drug discovery. However, its implementation and the adaptation of high-content assays to human embryonic stem cells (hESCs) have been hampered by multiple technical challenges. Here we present a strategy to adapt hESCs to HTS conditions, resulting in an assay suitable for the discovery of small molecules that drive hESC self-renewal or differentiation. Use of this new assay has led to the identification of several marketed drugs and natural compounds promoting short-term hESC maintenance and compounds directing early lineage choice during differentiation. Global gene expression analysis upon drug treatment defines known and novel pathways correlated to hESC self-renewal and differentiation. Our results demonstrate feasibility of hESC-based HTS and enhance the repertoire of chemical compounds for manipulating hESC fate. The availability of high-content assays should accelerate progress in basic and translational hESC biology.
Desbordes
Sabrina C
SC
Developmental Biology Program, Department of Neurosurgery, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA. sabrina.desbordes@crg.es
Placantonakis
Dimitris G
DG
Ciro
Anthony
A
Socci
Nicholas D
ND
Lee
Gabsang
G
Djaballah
Hakim
H
Studer
Lorenz
L
D000818:Animals; D001681:Biological Assay; D002454:Cell Differentiation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D033041:Embryo Research; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D006801:Humans; D051379:Mice; D046228:Microarray Analysis; D012038:Regeneration / Q000187:drug effects; D012107:Research Design / Q000639:trends*; D054852:Small Molecule Libraries / Q000494:pharmacology* / Q000627:therapeutic use
D054852:Small Molecule Libraries