Menzorov Aleksei G, Matveeva Natalia M, Markakis Marios N, Fishman Venyamin S, Christensen Knud, Khabarova Anna A, Pristyazhnyuk Inna E, Kizilova Elena A, Cirera Susanna, Anistoroaei Razvan, Serov Oleg L
BMC Genomics. 2015;16 Suppl 13(Suppl 13):S6. doi: 10.1186/1471-2164-16-S13-S6. Epub 2015 Dec 16.
Recently fibroblasts of many mammalian species have been reprogrammed to pluripotent state using overexpression of several transcription factors. This technology allows production of induced pluripotent stem (iPS) cells with properties similar to embryonic stem (ES) cells. The completeness of reprogramming process is well studied in such species as mouse and human but there is not enough data on other species. We produced American mink (Neovison vison) ES and iPS cells and compared these cells using transcriptome analysis.
We report the generation of 10 mink ES and 22 iPS cell lines. The majority of the analyzed cell lines had normal diploid chromosome number. The only ES cell line with XX chromosome set had both X-chromosomes in active state that is characteristic of pluripotent cells. The pluripotency of ES and iPS cell lines was confirmed by formation of teratomas with cell types representing all three germ layers. Transcriptome analysis of mink embryonic fibroblasts (EF), two ES and two iPS cell lines allowed us to identify 11831 assembled contigs which were annotated. These led to a number of 6891 unique genes. Of these 3201 were differentially expressed between mink EF and ES cells. We analyzed expression levels of these genes in iPS cell lines. This allowed us to show that 80% of genes were correctly reprogrammed in iPS cells, whereas approximately 6% had an intermediate expression pattern, about 7% were not reprogrammed and about 5% had a "novel" expression pattern. We observed expression of pluripotency marker genes such as Oct4, Sox2 and Rex1 in ES and iPS cell lines with notable exception of Nanog.
We had produced and characterized American mink ES and iPS cells. These cells were pluripotent by a number of criteria and iPS cells exhibited effective reprogramming. Interestingly, we had showed lack of Nanog expression and consider it as a species-specific feature.
最近,通过过表达几种转录因子,许多哺乳动物物种的成纤维细胞已被重编程为多能状态。这项技术使得诱导多能干细胞(iPS细胞)得以产生,其特性与胚胎干细胞(ES细胞)相似。在小鼠和人类等物种中,重编程过程的完整性已得到充分研究,但关于其他物种的数据还不够。我们制备了美国水貂(Neovison vison)的ES细胞和iPS细胞,并通过转录组分析对这些细胞进行了比较。
我们报告了10个水貂ES细胞系和22个iPS细胞系的产生。大多数分析的细胞系具有正常的二倍体染色体数目。唯一具有XX染色体组的ES细胞系的两条X染色体均处于活跃状态,这是多能细胞的特征。ES细胞系和iPS细胞系的多能性通过形成包含代表所有三个胚层的细胞类型的畸胎瘤得以证实。对水貂胚胎成纤维细胞(EF)、两个ES细胞系和两个iPS细胞系进行转录组分析,使我们能够鉴定出11831个已注释的组装重叠群。这些重叠群对应6891个独特基因。其中3201个基因在水貂EF细胞和ES细胞之间存在差异表达。我们分析了这些基因在iPS细胞系中的表达水平。这使我们能够表明,80%的基因在iPS细胞中被正确重编程,而约6%具有中间表达模式,约7%未被重编程,约5%具有“新”表达模式。我们在ES细胞系和iPS细胞系中观察到多能性标记基因如Oct4、Sox2和Rex1的表达,但Nanog是个明显的例外。
我们制备并鉴定了美国水貂的ES细胞和iPS细胞。这些细胞在多个标准下具有多能性,iPS细胞表现出有效的重编程。有趣的是,我们发现缺乏Nanog表达,并将其视为物种特异性特征。