Identification and characterization of small-molecule ligands that maintain pluripotency of human embryonic stem cells.
hESCs (human embryonic stem cells) offer great potential for pharmaceutical research and development and, potentially, for therapeutic use. However, improvements in cell culture are urgently required to allow the scalable production of large numbers of cells that maintain pluripotency. Supplementing feeder-free conditions with either EHNA [erythro-9-(2-hydroxy-3-nonyl)adenine] or readily synthesized analogues of this compound maintains hESC pluripotency in the absence of exogenous cytokines. When the hESC lines SA121 or SA461 were maintained in feeder-free conditions with EHNA they displayed no reduction in stem-cell-associated markers such as Nanog, Oct4 (octamer-binding protein 4) and SSEA4 (stage-specific embryonic antigen 4) when compared with cells maintained in full feeder-free conditions that included exogenously added bFGF (basic fibroblast growth factor). Spontaneous differentiation was reversibly suppressed by the addition of EHNA, but EHNA did not limit efficient spontaneous or directed differentiation following its removal. We conclude that EHNA or related compounds offers a viable alternative to exogenous cytokine addition in maintaining hESC cultures in a pluripotent state and might be a particularly useful replacement for bFGF for large-scale or GMP (good manufacturing practice)-compliant processes.
Burton
Peter
P
Neuroscience and Molecular Pharmacology, Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Adams
David R
DR
Abraham
Achamma
A
Allcock
Robert W
RW
Jiang
Zhong
Z
McCahill
Angela
A
Gilmour
Jane
J
McAbney
John
J
Kane
Nicole M
NM
Baillie
George S
GS
McKenzie
Fergus R
FR
Baker
Andrew H
AH
Houslay
Miles D
MD
Mountford
Joanne C
JC
Milligan
Graeme
G
D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D008024:Ligands; D039904:Pluripotent Stem Cells / Q000187:drug effects* / Q000502:physiology; D054852:Small Molecule Libraries / Q000032:analysis* / Q000494:pharmacology*
D008024:Ligands; D054852:Small Molecule Libraries