Transcriptional profiling of rhesus monkey embryonic stem cells
Transcriptional profiling of rhesus monkey embryonic stem cells
复制标题
DOI：
10.1095/biolreprod.106.053868
复制
发表时间：
2006-12-01
影响因子：
3.6
通讯作者：
Wolf, Don P.
中科院分区：
文献类型：
--
作者：
Byrne, James A.;Mitalipov, Shoukhrat M.;Wolf, Don P.
胚胎干细胞(ESCs)可能能够治愈或缓解各种退行性疾病的症状。然而，关于存活、功能和肿瘤形成的悬而未决的问题意味着应该采取谨慎的方法来推动ESCs进入人类临床试验。恒河猴为制定预防免疫排斥反应的策略和测试基于胚胎干细胞的医疗治疗的可行性、安全性和有效性提供了理想的模式生物。猕猴胚胎干细胞的转录图谱为该物种的临床前胚胎干细胞研究提供了基础。在本研究中，我们利用基因芯片技术、免疫细胞化学、逆转录-聚合酶链式反应(RT-PCR)和实时定量聚合酶链式反应(QPCR)对恒河猴胚胎干细胞进行了鉴定和转录分析。我们鉴定了367个在五个不同的ESC系中高度保守(>；85%)的STRNISH基因候选。恒河猴ESC系在广泛的Pou5f1(也称为OCT4)表达水平上保持着多潜能未分化状态，并且恒河猴、小鼠和人类的严厉基因之间的比较揭示了五个哺乳动物的严厉基因：CCNB1、GDF3、LEFTB、Pou5f1和NANOG。这五个哺乳动物基因在恒河猴、小鼠和人类胚胎干细胞中强烈表达，尽管只是在未分化状态下表达，并代表了哺乳动物的核心关键严厉因子。
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 preclinical 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 sternness 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 sternness genes revealed five mammalian sternness 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 sternness factors.