Motedayyen Hossein, Esmaeil Nafiseh, Tajik Nader, Khadem Fahimeh, Ghotloo Somayeh, Khani Behnaz, Rezaei Abbas
Department of Immunology, Faculty of Medicine, Isfahan University of Medical Sciences, Hezar-Jereeb Ave, Isfahan, Iran.
Immunology Research Center (IRC), Iran University of Medical Sciences, Tehran, Iran.
BMC Res Notes. 2017 Nov 2;10(1):552. doi: 10.1186/s13104-017-2880-6.
Human amniotic epithelial cells (hAECs) which are isolated from the amniotic membrane have stem cell-like properties and immunomodulatory effects. Several protocols have been proposed for isolation of hAECs, nevertheless, there is no report concerning isolation of highly viable hAECs, with desirable yield, and without significant purity reduction. In the current study, a detailed protocol with some modification of previous ones is presented in which the amendments led to isolation of hAECs with high purity, yield and viability. Moreover, isolated hAECs were subjected to immuno-phenotyping and their physiological status was assessed using a proliferation assay.
The average yield of obtained hAECs using the new modified method was 190 × 10 cells with a mean viability of 87%, with less than 1% contamination with mesenchymal stem cells (MSCs). The isolated cells were > 95% positive for the epithelial cell markers. The lowest initial plating efficiency of the cells was 80%. Freshly isolated hAECs had the ability to proliferate for 5-6 passages in a standard culture medium.
从羊膜分离出的人羊膜上皮细胞（hAECs）具有干细胞样特性和免疫调节作用。已经提出了几种分离hAECs的方案，然而，尚无关于分离出高活力、产量理想且纯度无显著降低的hAECs的报道。在本研究中，我们提出了一个对先前方案进行了一些修改的详细方案，这些修改使得能够分离出高纯度、高产量和高活力的hAECs。此外，对分离出的hAECs进行免疫表型分析，并使用增殖试验评估其生理状态。
使用新的改良方法获得的hAECs的平均产量为190×10个细胞，平均活力为87%，间充质干细胞（MSCs）污染率低于1%。分离出的细胞上皮细胞标志物阳性率>95%。细胞的最低初始接种效率为80%。新鲜分离的hAECs能够在标准培养基中增殖5 - 6代。
