Establishment and characterization of immortalized human amniotic epithelial cells.
Establishment and characterization of immortalized human amniotic epithelial cells.
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DOI：
10.1089/cell.2012.0021
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发表时间：
2013-02
影响因子：
1.6
通讯作者：
Kaixuan Zhou;C. Koike;Toshiko Yoshida;M. Okabe;M. Fathy;S. Kyo;T. Kiyono;S. Saito;T. Nikaido
中科院分区：
文献类型：
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作者：
Kaixuan Zhou;C. Koike;Toshiko Yoshida;M. Okabe;M. Fathy;S. Kyo;T. Kiyono;S. Saito;T. Nikaido
人羊膜上皮细胞（HAE）具有低免疫原性，并具有有效的免疫抑制特性。HAE也具有与干细胞相似的几个特征，并且它们在分娩后被丢弃。因此，它们可以潜在地用于细胞治疗，伦理问题较少。由于羊膜上皮细胞的寿命较短，因此我们的目标是建立和鉴定永生化的人羊膜上皮细胞（iHAE）。将HAE与病毒致癌基因E6/E7和人端粒酶逆转录酶（hTERT）一起引入以产生iHAE。这些iHAE已增殖约200个群体倍增（PD）至少12个月。免疫组化和逆转录聚合酶链反应（RT-PCR）检测干细胞标志物（Oct 3/4、Nanog、Sox 2、Klf 4）和上皮标志物（CK 5、CK 18）的高表达。这些iHAE在超低附着培养皿中扩增以形成类似于上皮干细胞/前体细胞的球状体。流式细胞术检测间充质干细胞标志物（CD 44、CD 73、CD 90、CD 105）和体细胞标志物（CD 24、CD 29、CD 271、Nestin）的高表达。iHAE显示成脂、成骨、神经元和心脏分化能力。总之，已经建立了具有干细胞特征的HAE的永生化，使得这些iHAE可用于细胞治疗和再生医学。
Human amniotic epithelial cells (HAEs) have a low immunogenic profile and possess potent immunosuppressive properties. HAEs also have several characteristics similar to stem cells, and they are discarded after parturition. Thus, they could potentially be used in cell therapy with fewer ethical problems. HAEs have a short life, so our aim is to establish and characterize immortalized human amniotic epithelial cells (iHAEs). HAEs were introduced with viral oncogenes E6/E7 and with human telomerase reverse transcriptase (hTERT) to create iHAEs. These iHAEs have proliferated around 200 population doublings (PDs) for at least 12 months. High expression of stem cell markers (Oct 3/4, Nanog, Sox2, Klf4) and epithelial markers (CK5, CK18) were detected by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR). These iHAEs were expanded in ultra-low-attachment dishes to form spheroids similarly to epithelial stem/precursor cells. High expression of mesenchymal (CD44, CD73, CD90, CD105) and somatic (CD24, CD29, CD271, Nestin) stem cell markers was detected by flow cytometry. The iHAEs showed adipogenic, osteogenic, neuronal, and cardiac differentiation abilities. In conclusion, the immortalization of HAEs with the characteristics of stem cells has been established, allowing these iHAEs to become useful for cell therapy and regenerative medicine.