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Transcriptional Profiling of Rhesus Monkey Embryonic Stem Cells¹

Oregon National Primate Research Center,³ Oregon Health & Science University, Beaverton, Oregon 97006 Departments of Obstetrics and Gynecology⁴ and Physiology and Pharmacology,⁵ Oregon Health & Science University, Portland, Oregon 97201

ABSTRACT

Embryonic stem cells (ESCs) may be able to cure or alleviate the symptoms of various degenerative diseases. However, unresolved issues regarding survival, functionality, and tumor formation mean a prudent approach should be adopted towards advancing ESCs into human clinical trials. The rhesus monkey provides an ideal model organism for developing strategies to prevent immune rejection and test the feasibility, safety, and efficacy of ESC-based medical treatments. Transcriptional profiling of rhesus monkey ESCs provides a foundation for pre-clinical ESC research in this species. In the present study, we used microarray technology, immunocytochemistry, reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR (qPCR) to characterize and transcriptionally profile rhesus monkey ESCs. We identified 367 stemness gene candidates that were highly (>85%) conserved across five different ESC lines. Rhesus monkey ESC lines maintained a pluripotent undifferentiated state over a wide range of POU5F1 (also known as OCT4) expression levels, and comparisons between rhesus monkey, mouse, and human stemness genes revealed five mammalian stemness genes: CCNB1, GDF3, LEFTB, POU5F1, and NANOG. These five mammalian genes are strongly expressed in rhesus monkey, mouse, and human ESCs, albeit only in the undifferentiated state, and represent the core key mammalian stemness factors.

embryo, gene regulation, microarray, rhesus, stem cells

¹Supported by Oregon National Primate Research Center pilot grant GPRC46174 (to J.A.B.) and National Institute of Health grant RR00163 (to P.K.).

Correspondence: ² FAX: 503 690 5563; e-mail: wolfd{at}ohsu.edu

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