Ginis Irene, Luo Yongquan, Miura Takumi, Thies Scott, Brandenberger Ralph, Gerecht-Nir Sharon, Amit Michal, Hoke Ahmet, Carpenter Melissa K, Itskovitz-Eldor Joseph, Rao Mahendra S
Stem Cell Section, Laboratory of Neurosciences, National Institute on Aging, NIH, Baltimore, MD 21224, USA.
Dev Biol. 2004 May 15;269(2):360-80. doi: 10.1016/j.ydbio.2003.12.034.
We compared gene expression profiles of mouse and human ES cells by immunocytochemistry, RT-PCR, and membrane-based focused cDNA array analysis. Several markers that in concert could distinguish undifferentiated ES cells from their differentiated progeny were identified. These included known markers such as SSEA antigens, OCT3/4, SOX-2, REX-1 and TERT, as well as additional markers such as UTF-1, TRF1, TRF2, connexin43, and connexin45, FGFR-4, ABCG-2, and Glut-1. A set of negative markers that confirm the absence of differentiation was also developed. These include genes characteristic of trophoectoderm, markers of germ layers, and of more specialized progenitor cells. While the expression of many of the markers was similar in mouse and human cells, significant differences were found in the expression of vimentin, beta-III tubulin, alpha-fetoprotein, eomesodermin, HEB, ARNT, and FoxD3 as well as in the expression of the LIF receptor complex LIFR/IL6ST (gp130). Profound differences in cell cycle regulation, control of apoptosis, and cytokine expression were uncovered using focused microarrays. The profile of gene expression observed in H1 cells was similar to that of two other human ES cell lines tested (line I-6 and clonal line-H9.2) and to feeder-free subclones of H1, H7, and H9, indicating that the observed differences between human and mouse ES cells were species-specific rather than arising from differences in culture conditions.
我们通过免疫细胞化学、逆转录聚合酶链反应(RT-PCR)以及基于膜的聚焦cDNA阵列分析,比较了小鼠和人类胚胎干细胞的基因表达谱。确定了几个协同作用可将未分化的胚胎干细胞与其分化后代区分开来的标志物。这些标志物包括已知的标志物,如阶段特异性胚胎抗原(SSEA)、八聚体结合转录因子3/4(OCT3/4)、SRY-盒转录因子2(SOX-2)、Rex-1蛋白和端粒酶逆转录酶(TERT),以及其他标志物,如未分化胚胎干细胞转录因子1(UTF-1)、端粒重复结合因子1(TRF1)、端粒重复结合因子2(TRF2)、连接蛋白43、连接蛋白45、成纤维细胞生长因子受体4(FGFR-4)、ATP结合盒转运体G2(ABCG-2)和葡萄糖转运蛋白1(Glut-1)。还开发了一组确认无分化的阴性标志物。这些标志物包括滋养外胚层特征性基因、胚层标志物以及更特化的祖细胞标志物。虽然许多标志物在小鼠和人类细胞中的表达相似,但在波形蛋白、β-III微管蛋白、甲胎蛋白、中胚层决定因子、HEB、芳香烃受体核转运蛋白(ARNT)和叉头框蛋白D3(FoxD3)的表达以及白血病抑制因子受体复合物LIFR/IL6ST(gp130)的表达方面发现了显著差异。使用聚焦微阵列揭示了细胞周期调控、细胞凋亡控制和细胞因子表达方面的深刻差异。在H1细胞中观察到的基因表达谱与另外两个测试的人类胚胎干细胞系(I-6系和克隆系-H9.2)以及H1、H7和H9的无饲养层亚克隆相似,表明观察到的人类和小鼠胚胎干细胞之间的差异是物种特异性的,而非源于培养条件的差异。