Wu Xulun, Gao Fan, Wu Yangnan, Sun Ruiyang, Guan Weijun, Tian Xiuzhi
Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing, 100193, People's Republic of China.
College of Human Movement Science, Harbin Sport University, Harbin, 150040, Heilongjiang, People's Republic of China.
Cytotechnology. 2019 Apr;71(2):539-551. doi: 10.1007/s10616-019-00299-1. Epub 2019 Feb 28.
Amniotic epithelial cells (AECs), isolated from placenta, have epithelial cells and stem cells characteristics. Most of the previous studies focused on the biological characteristics of human amniotic epithelial cells, which demonstrated amniotic epithelial cells not only had low immunogenicity and potent potential to differentiate into three germ layers, but also could secrete various immunomodulatory factors. However, compared to study on human amniotic epithelial cells, few studies have been done on other animals. In this study, sheep amniotic epithelial cells were successfully isolated and their surface makers were accessed by immunofluorescence assay, and found that AECs were expressed Oct4 and Sox2, which were necessary for maintaining the undifferentiated state of pluripotent stem cells. Based on cloning efficiency and growth kinetics assay, AECs were found to possess self-renewal capacity and the growth curve was S-shaped. In addition, AECs could be induced into adipocytes, osteoblasts and chondrocytes in vitro, showing they had multi-differential ability. Reverse transcription-polymerase chain reaction results showed that AECs expressed CD29, CD44, CD90 and CK19, and didn't expressed CD34, CD45 and the telomerase gene (TERT). Little change in chromosome number was observed in AEC cultures for up to at least the first ten passages. In summary, this study results revealed that sheep AECs possessed more advantages for cell therapy and might play a key role in cell therapy and regenerative medicine in the future.
从胎盘中分离出的羊膜上皮细胞（AECs）具有上皮细胞和干细胞的特征。此前的大多数研究集中在人羊膜上皮细胞的生物学特性上，这些研究表明羊膜上皮细胞不仅免疫原性低，具有分化为三个胚层的强大潜力，而且还能分泌多种免疫调节因子。然而，与对人羊膜上皮细胞的研究相比，对其他动物的研究较少。在本研究中，成功分离出绵羊羊膜上皮细胞，并通过免疫荧光测定法检测其表面标志物，发现AECs表达Oct4和Sox2，这是维持多能干细胞未分化状态所必需的。基于克隆效率和生长动力学分析，发现AECs具有自我更新能力，生长曲线呈S形。此外，AECs在体外可被诱导分化为脂肪细胞、成骨细胞和软骨细胞，表明它们具有多向分化能力。逆转录-聚合酶链反应结果显示，AECs表达CD29、CD44、CD90和CK19，不表达CD34、CD45和端粒酶基因（TERT）。在至少前十代的AEC培养物中，未观察到染色体数量有明显变化。总之，本研究结果表明绵羊AECs在细胞治疗方面具有更多优势，可能在未来的细胞治疗和再生医学中发挥关键作用。
