König Julia, Lang Ingrid, Siwetz Monika, Fröhlich Julia, Huppertz Berthold
Institute of Cell Biology, Histology and Embryology, Medical University of Graz, Harrachgasse 21/7, 8010, Graz, Austria.
Cell Tissue Bank. 2014 Jun;15(2):193-8. doi: 10.1007/s10561-013-9415-8. Epub 2013 Dec 11.
The amnionic membrane is a rich source of multipotent mesenchymal stromal cells (hAMSC), which are readily available and show a potential use in regenerative medicine and tissue engineering. Before these cells can be applied clinically, careful characterization is necessary, especially as primary cells are known to change their phenotype in culture. We analyzed the mesenchymal phenotype of hAMSC at different stages after isolation using immunohistochemistry. Shortly after isolation (1 day), 92 % (± 7 %) of the hAMSC expressed the mesenchymal marker vimentin, 2 % (± 1 %) stained for the epithelial marker cytokeratin-7 and 5 % (± 4 %) co-expressed these markers. After 5 days, the double positive cells slightly increased to 7 % (± 3 %), while exclusive expression of cytokeratin-7 or vimentin remained unchanged (1 % ± 2 % and 92 % ± 1 %, respectively). After the first passage, all attached cells were vimentin-positive, while 54 % (± 9 %) co-expressed cytokeratin-7 and vimentin. Thus, we conclude that under culture, hAMSC adopt a hybrid mesenchymal-epithelial phenotype. It is also essential to perform microscopical examination during the first days after isolation to detect contaminations with human amnion-derived epithelial cells in cultures of hAMSC.
羊膜是多能间充质基质细胞（hAMSC）的丰富来源，这些细胞易于获取，在再生医学和组织工程中显示出潜在用途。在这些细胞能够临床应用之前，进行仔细的特性鉴定是必要的，尤其是因为已知原代细胞在培养过程中会改变其表型。我们使用免疫组织化学分析了分离后不同阶段hAMSC的间充质表型。分离后不久（1天），92%（±7%）的hAMSC表达间充质标志物波形蛋白，2%（±1%）的细胞被上皮标志物细胞角蛋白-7染色，5%（±4%）的细胞共表达这些标志物。5天后，双阳性细胞略有增加至7%（±3%），而细胞角蛋白-7或波形蛋白的单独表达保持不变（分别为1%±2%和92%±1%）。首次传代后，所有贴壁细胞波形蛋白均为阳性，而54%（±9%）的细胞共表达细胞角蛋白-7和波形蛋白。因此，我们得出结论，在培养条件下，hAMSC呈现出混合的间充质-上皮表型。在分离后的头几天进行显微镜检查以检测hAMSC培养物中是否有人羊膜来源的上皮细胞污染也很重要。
