Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential
Molecular and phenotypic characterization of human amniotic fluid cells and their differentiation potential
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DOI：
10.1038/sj.cr.7310043
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发表时间：
2006-04-01
期刊：
影响因子：
44.1
通讯作者：
Lazzari, Lorenza
中科院分区：
文献类型：
--
作者：
Bossolasco, Patrizia;Montemurro, Tiziana;Lazzari, Lorenza
这项研究的主要目标是确定一种新的人类多能细胞来源，克服胚胎干细胞研究中涉及的伦理问题和大多数成体干细胞的有限可用性。羊水细胞（AFC）是常规获得产前诊断，并可以在体外扩增，但目前的知识，其起源和性质是有限的。将20个AFC样品在培养中暴露于成脂、成骨、成神经和成肌培养基。采用免疫细胞化学、RT-PCR和Western blotting检测分化情况。治疗前，AFC表现出不均匀的形态。RT-PCR检测结果显示，MyoD、Myf-5、MRF 4、Myogenin和Desmin呈阴性，但骨钙素、PPARgamma 2、GAP 43、NSE、Nestin、MAP 2、GFAP和β微管蛋白III呈阳性。这些细胞表达Oct-4、雷克斯-1和Runx-1，它们是未分化干细胞状态的特征。通过免疫细胞化学，他们表达神经胶质蛋白，间充质和上皮标记。培养后，当主要细胞成分为成纤维细胞时，AFC分化为脂肪细胞和成骨细胞。早期和晚期神经元抗原仍然存在2周后，在神经特异性培养基中的培养，即使没有神经元形态检测。我们的研究结果提供的证据表明，人羊水中含有祖细胞与多谱系的潜力，显示干细胞和组织特异性基因/蛋白存在的几个谱系。
The main goal of the study was to identify a novel source of human multipotent cells, overcoming ethical issues involved in embryonic stem cell research and the limited availability of most adult stem cells. Amniotic fluid cells (AFCs) are routinely obtained for prenatal diagnosis and can be expanded in vitro; nevertheless current knowledge about their origin and properties is limited. Twenty samples of AFCs were exposed in culture to adipogenic, osteogenic, neurogenic and myogenic media. Differentiation was evaluated using immunocytochemistry, RT-PCR and Western blotting. Before treatments, AFCs showed heterogeneous morphologies. They were negative for MyoD, Myf-5, MRF4, Myogenin and Desmin but positive for osteocalcin, PPARgamma2, GAP43, NSE, Nestin, MAP2, GFAP and beta tubulin III by RT-PCR. The cells expressed Oct-4, Rex-1 and Runx-1, which characterize the undifferentiated stem cell state. By immunocytochemistry they expressed neural-glial proteins, mesenchymal and epithelial markers. After culture, AFCs differentiated into adipocytes and osteoblasts when the predominant cellular component was fibroblastic. Early and late neuronal antigens were still present after 2 week culture in neural specific media even if no neuronal morphologies were detectable. Our results provide evidence that human amniotic fluid contains progenitor cells with multi-lineage potential showing stem and tissue-specific gene/protein presence for several lineages.