Adipocyte differentiation in human embryonic stem cells transduced with Oct4 shRNA lentivirus

Hannan, Nicholas R.F. and Wolvetang, Ernst J. (2009) Adipocyte differentiation in human embryonic stem cells transduced with Oct4 shRNA lentivirus. Stem Cells Development, 18 4: 653-660.


Author Hannan, Nicholas R.F.
Wolvetang, Ernst J.
Title Adipocyte differentiation in human embryonic stem cells transduced with Oct4 shRNA lentivirus
Journal name Stem Cells Development   Check publisher's open access policy
ISSN 1547-3287
Publication date 2009-05-11
Year available 2008
Sub-type Article (original research)
DOI 10.1089/scd.2008.0160
Volume 18
Issue 4
Start page 653
End page 660
Total pages 8
Editor D. English
Place of publication New Rochelle, NY, USA
Publisher Mary Ann Liebert Publishers
Collection year 2009
Language eng
Subject C1
100404 Regenerative Medicine (incl. Stem Cells and Tissue Engineering)
060103 Cell Development, Proliferation and Death
970106 Expanding Knowledge in the Biological Sciences
Abstract Oct4 is one of the master pluripotency genes that controls differentiation of human embryonic stem cells (hESCs). We generated HES2 and HES3 hESC lines stably transduced with lentivirus carrying Oct4 short hairpin RNA (shRNA) that display 80–90% reduction of Oct4 expression. Analysis of pluripotency marker expression shows that these Oct4 shRNA-transduced hESCs display normal wild-type expression levels of the pluripotency marker CD9 but an absence of GCTM2 expression. These hESC-derived adipocyte precursor cells display a characteristic morphology and can be propagated and cryopreserved as a standard stem cell line. Interestingly, Oct4 shRNA-transduced hESCs display a remarkably high lineage-specific spontaneous differentiation toward adipocytes. After two weeks of spontaneous differentiation under feeder-free conditions, 60–70% of cells display a mature adipocyte morphology as well as the expression of multiple adipocyte-specific mRNAs as assessed by RT-PCR. The upregulation of trophoblast, mesoderm, and endoderm transcripts is, however, also detected in these spontaneously differentiating cultures. These Oct4 shRNA hESCs will be an interesting model system to study human fetal adipogenesis and constitutes a renewable resource for obesity drug screening purposes.
Keyword Transcription Factor
Mammalian Embryo
Self-renewal
Pluripotency
Commitment
Generation
Derivation
networks
Lineage
BMP-4
Q-Index Code C1
Q-Index Status Confirmed Code
Additional Notes Online Ahead of Editing: September 2, 2008

Document type: Journal Article
Sub-type: Article (original research)
Collections: 2009 Higher Education Research Data Collection
Australian Institute for Bioengineering and Nanotechnology Publications
 
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