Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) blocks differentiation and maintains the expression of pluripotency markers in human embryonic stem cells

Autor: Robert W. Allcock, John P. McAbney, Zhong Jiang, Nicole M. Kane, David R. Adams, Graeme Milligan, Joanne C. Mountford, Peter Burton, Alexandra Kaupisch, Achamma Abraham, Jane Gilmour, Angela McCahill, Miles D. Houslay, Andrew H. Baker, George S. Baillie
Přispěvatelé: Faculty of Biomedical and Life Sciences, University of Glasgow
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Homeobox protein NANOG
Pluripotent Stem Cells
Stage-Specific Embryonic Antigens
Time Factors
Phosphodiesterase Inhibitors
Cellular differentiation
Basic fibroblast growth factor
Cell Culture Techniques
Biology
Biochemistry
Second Messenger Systems
Cell Line
03 medical and health sciences
chemistry.chemical_compound
QH345
Structure-Activity Relationship
0302 clinical medicine
Directed differentiation
medicine
Adenosine Deaminase Inhibitors
Humans
Induced pluripotent stem cell
Molecular Biology
Embryonic Stem Cells
030304 developmental biology
Homeodomain Proteins
Neurons
0303 health sciences
Adenine
Gene Expression Profiling
Gene Expression Regulation
Developmental
Life Sciences
Cell Differentiation
Cell Biology
Nanog Homeobox Protein
Molecular biology
Antigens
Differentiation
chemistry
Cell culture
embryonic structures
Phosphodiesterase 2
EHNA
Hydrophobic and Hydrophilic Interactions
Octamer Transcription Factor-3
030217 neurology & neurosurgery
medicine.drug
Zdroj: Biochemical Journal
Biochemical Journal, Portland Press, 2010, 432 (3), pp.575-584. ⟨10.1042/BJ20100726⟩
ISSN: 0264-6021
1470-8728
DOI: 10.1042/BJ20100726⟩
Popis: hESCs (human embryonic stem cells) have enormous potential for use in pharmaceutical development and therapeutics; however, to realize this potential, there is a requirement for simple and reproducible cell culture methods that provide adequate numbers of cells of suitable quality. We have discovered a novel way of blocking the spontaneous differentiation of hESCs in the absence of exogenous cytokines by supplementing feeder-free conditions with EHNA [erythro-9-(2-hydroxy-3-nonyl)adenine], an established inhibitor of ADA (adenosine deaminase) and cyclic nucleotide PDE2 (phosphodiesterase 2). hESCs maintained in feeder-free conditions with EHNA for more than ten passages showed no reduction in hESC-associated markers including NANOG, POU5F1 (POU domain class 5 transcription factor 1, also known as Oct-4) and SSEA4 (stage-specific embryonic antigen 4) compared with cells maintained in feeder-free conditions containing bFGF (basic fibroblast growth factor). Spontaneous differentiation was reversibly suppressed by the addition of EHNA, but, upon removing EHNA, hESC populations underwent efficient spontaneous, multi-lineage and directed differentiation. EHNA also acts as a strong blocker of directed neuronal differentiation. Chemically distinct inhibitors of ADA and PDE2 lacked the capacity of EHNA to suppress hESC differentiation, suggesting that the effect is not driven by inhibition of either ADA or PDE2. Preliminary structure–activity relationship analysis found the differentiation-blocking properties of EHNA to reside in a pharmacophore comprising a close adenine mimetic with an extended hydrophobic substituent in the 8- or 9-position. We conclude that EHNA and simple 9-alkyladenines can block directed neuronal and spontaneous differentiation in the absence of exogenous cytokine addition, and may provide a useful replacement for bFGF in large-scale or cGMP-compliant processes.
Databáze: OpenAIRE
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