title: Establishment and characterization of immortalized human amniotic epithelial cells.
abstract: 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.
authors.affiliation: Department of Regenerative Medicine, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Japan.
authors.forename: Kaixuan
authors.initials: K
authors.lastname: Zhou
authors.forename: Chika
authors.initials: C
authors.lastname: Koike
authors.forename: Toshiko
authors.initials: T
authors.lastname: Yoshida
authors.forename: Motonori
authors.initials: M
authors.lastname: Okabe
authors.forename: Moustafa
authors.initials: M
authors.lastname: Fathy
authors.forename: Satoru
authors.initials: S
authors.lastname: Kyo
authors.forename: Tohru
authors.initials: T
authors.lastname: Kiyono
authors.forename: Shigeru
authors.initials: S
authors.lastname: Saito
authors.forename: Toshio
authors.initials: T
authors.lastname: Nikaido
mesh_terms: D000650:Amnion / Q000166:cytology* / Q000378:metabolism*; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D002461:Cell Line, Transformed / Q000166:cytology* / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D000090062:Kruppel-Like Factor 4; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis / Q000235:genetics; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics
chemical_list: D015703:Antigens, CD; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D050725:Papillomavirus E7 Proteins; D014157:Transcription Factors; C509186:TERT protein, human; D019098:Telomerase