由有胚胎的干细胞的熔化的 Reprogramming 体的房间不在老鼠引起 Dlk1-Dio3 区域的 silencing

• 作者: Nariman R Battulin Anna A Khabarova Ul’yana A Boyarskikh Aleksey G Menzorov Maxim L Filipenko Oleg L Serov
• 来源期刊: 世界干细胞杂志：英文版（电子版）
• 学科: 生物学
• 关键词: Cell FUSION EMBRYONIC stem cells REPROGRAMMING PLURIPOTENCY Imprinted Dlk1-Dio3 region DNA METHYLATION
• 年，卷（期）: 2012,（8）
• 页码范围: 87-93
• 页数: 7页
• 分类号: Q813

摘要

AIM:To examine the imprinted Dlk1-Dio3 locus in pluripotent embryonic stem(ES)cell/fibroblast hybrid cells.METHODS:Gtl2,Rian,and Mirg mRNA expression in mouse pluripotent ES cell/fibroblast hybrid cells was examined by real-time reverse transcription-polymerase chain reaction.Pyrosequencing and bisulfate sequencing were used to determine the DNA methylation level of the Dlk1-Dio3 locus imprinting control region. RESULTS:The selected hybrid clones had a near-tetraploid karyotype and were highly pluripotent judging from their capacity to generate chimeric embryos and adult chimeras.Our data clearly demonstrate that Gtl2,Rian,and Mirg,which are imprinted genes within the Dlk1-Dio3 locus,are active in all examined ES cell/fibroblast hybrid clones.In spite of interclonal variability,the expression of the imprinted genes is comparable to that of ES cells and fibroblasts.Quantitative analysis of the DNA methylation status of the intergenic differentially methylated region(IG DMR)within the Dlk1-Dio3 locus by pyrosequencing and bisulfite sequencing clearly showed that the DNA methylation status of the imprinted region in the tested hybrid clones was comparable to that of both ES cells and fibroblasts.CONCLUSION:Reprogramming process in a hybrid cell system is achieved without marked alteration of the imprinted Dlk1-Dio3 locus.