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- Title
Expression of Dppa2 during the differentiation of human embryonic stem cells.
- Authors
Chen, T. J.; Du, J.; Lu, G. X.
- Abstract
Introduction: Human embryonic stem cells (human ESC) are pluripotent cells derived from inner cell mass (ICM) of early blastocysts and have the ability to form virtually all cell types. This property holds great promise for the field of regenerative medicine. In recent years, some key regulators have identified essentially for sustaining the pluripotency of ESC, such as Oct4, Sox2 and Nanog. Dppa2 is a novel gene expressed specifically in ESCs. To explore its role in maintaining the pluripotency of ESC, we analysed its expression levels during differentiation of human ESC induced by all trans-retinoic acid (atRA) treatment and embryoid body (EB) formation. Materials/Methods: Undifferentiated human ESC were induced differentiation by atRA (10-6 M), and EB were cultured in EB medium. The undifferentiated human ESC were collected at day 0, differentiated human ESC were collected at day 4 and day 7, and EB were collected at day 5 and day 21. Total RNA was isolated using Trizol reagent (Invitrogen), cDNA was synthesized from 750 ng of DNase I (NEB)-treated total RNA in a 20 µl reaction with oligo dT18 primer using the RevertAid First Strand cDNA Synthesis Kit (Fermentas), endogenous gene expression levels were then measured by real-time PCR analysis with SYBR Green Supermix (Bio-Rad) using the iCycler System (Bio-Rad). Results: The expression of Dppa2 was reduced by 20% at day 4 and as much as 90% at day 7 under atRA treatment, while Oct4 and Nanog 43% and 70% at day 4, 93% and 97% at day 7, respectively. During EB formation, the expression of Dppa2 was reduced by 52% at day 5 and 73% at day 21, while Oct4 and Nanog 53% and 74% at day 5, both 99% at day 21. Conclusions: The similar expression pattern between Dppa2 and Oct4, Nanog indicated that this novel gene might take part in the regulation of the pluripotency by interaction with Oct4 and Nanog and it might be essential for maintaining the pluripotency of ESC.
- Subjects
EMBRYONIC stem cells; BLASTOCYST
- Publication
Reproductive BioMedicine Online (Reproductive Healthcare Limited), 2008, Vol 16, Issue S2, pS-33
- ISSN
1472-6483
- Publication type
Abstract
- DOI
10.1016/S1472-6483(10)61546-5