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Fritsch, Michael Kevin; Alexanian, Ruben; Singer, Don B.; Murdoch, Fern E.

Altered retinoic acid (RA)‐induced lineage specification of mouse embryonic stem cells (mESCs) following a transient pulse of a histone deacetylase inhibitor

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  • Medientyp: E-Artikel
  • Titel: Altered retinoic acid (RA)‐induced lineage specification of mouse embryonic stem cells (mESCs) following a transient pulse of a histone deacetylase inhibitor
  • Beteiligte: Fritsch, Michael Kevin; Alexanian, Ruben; Singer, Don B.; Murdoch, Fern E.
  • Erschienen: Wiley, 2009
  • Erschienen in: The FASEB Journal
  • Sprache: Englisch
  • DOI: 10.1096/fasebj.23.1_supplement.71.1
  • ISSN: 0892-6638; 1530-6860
  • Schlagwörter: Genetics ; Molecular Biology ; Biochemistry ; Biotechnology
  • Entstehung:
  • Anmerkungen:
  • Beschreibung: <jats:sec><jats:label /><jats:p>RA treatment during mESC differentiation results in predominantly neural specification. A transient pulse of the histone deacetylase inhibitor, Trichostatin A (TSA), during early mESC differentiation via embryoid body (EB) formation results in maintenance of undifferentiated characteristics. Our objective was to determine whether a pulse of TSA followed by RA alters neural lineage specification. Undifferentiated mESCs are exposed to 100 nM of TSA during the first 24h of 3 days of EB formation. EBs are plated and RA added for an additional 4 days. Selected RT‐PCR shows repressed neural marker expression and only a small increase in a few endoderm markers. Gene expression array studies analyzed using a lineage annotated gene database demonstrates a marked increase in expression of numerous placental genes and confirms diminished neural gene expression. Immunofluorescence for the trophoblast marker Plac1, shows increased levels in TSA‐pulsed, RA‐treated cells with minimal expression in RA‐only differentiated cells. Conversely, the neural specific product, Doublecortin, is highly expressed in the RA‐only treated cells and shows diminished protein levels in TSA‐pulsed, RA‐treated cells. We conclude that transient TSA treatment prior to RA induces a dramatic lineage switch characterized by repressed neural differentiation and marked trophoblast induction. Supported by NIH grant RO1DK064243 to MKF.</jats:p></jats:sec>