Oct-4-expressing cells in human amniotic fluid:: a new source for stem cell research?
Oct-4-expressing cells in human amniotic fluid:: a new source for stem cell research?
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DOI：
10.1093/humrep/deg279
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发表时间：
2003-07-01
影响因子：
6.1
通讯作者：
Hengstschläger, M
中科院分区：
文献类型：
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作者：
Prusa, AR;Marton, E;Hengstschläger, M
背景技术背景：研究人员和患者都希望，在未来，多能人类干细胞的分离将允许建立各种疾病的治疗概念。这方面的一个主要目标是确定多能干细胞的新来源。Oct-4是迄今已知在胚胎癌细胞、胚胎干细胞和胚胎生殖细胞中表达的多能人类干细胞的标志物。方法：通过RT-PCR分析来自人羊水样品的细胞的Oct-4、干细胞因子、波形蛋白和碱性磷酸酶的mRNA表达。Western blot和免疫细胞化学检测Oct-4蛋白表达。Oct-4阳性细胞也通过双重免疫染色分析细胞周期蛋白A蛋白的表达。研究结果：进行RT-PCR，Western印迹和免疫细胞化学分析表明，在人羊水中的背景下的Oct-4阴性细胞的一个独特的群体的细胞可以被发现，表达Oct-4在细胞核中。Oct-4阳性羊水细胞样本也表达干细胞因子、波形蛋白和碱性磷酸酶mRNA。Oct-4阳性羊水细胞分裂活跃，通过检测细胞周期蛋白A表达证实。结论：这里提出的结果表明，人羊水可能代表了一个新的来源，用于分离人Oct-4阳性干细胞，而不会引起与人类胚胎研究相关的伦理问题。
BACKGROUND: It is the hope of investigators and patients alike that in future the isolation of pluripotent human stem cells will allow the establishment of therapeutic concepts for a wide variety of diseases. A major aim in this respect is the identification of new sources for pluripotent stem cells. Oct-4 is a marker for pluripotent human stem cells so far known to be expressed in embryonal carcinoma cells, embryonic stem cells and embryonic germ cells. METHODS: Cells from human amniotic fluid samples were analysed for mRNA expression of Oct-4, stem cell factor, vimentin and alkaline phosphatase via RT-PCR. Oct-4 protein expression was investigated by Western blot analysis and immunocytochemistry. Oct-4-positive cells were also analysed for the expression of cyclin A protein via double immunostaining. RESULTS: Performing RT-PCR, Western blot and immunocytochemical analyses revealed that in human amniotic fluid in the background of Oct-4-negative cells a distinct population of cells can be found, which express Oct-4 in the nucleus. Oct-4-positive amniotic fluid cell samples also express stem cell factor, vimentin and alkaline phosphatase mRNA. The Oct-4-positive amniotic fluid cells are actively dividing, proven by the detection of cyclin A expression. CONCLUSIONS: The results presented here suggest that human amniotic fluid may represent a new source for the isolation of human Oct-4-positive stem cells without raising the ethical concerns associated with human embryonic research.