Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration |
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Authors: | Simat Siti Fatimah Kienhui Chua Geok Chin Tan Tengku Ibrahim Azmi Ay Eeng Tan Hayati Abdul Rahman |
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Affiliation: | 1. Department of Developmental and Cell Biology, Sue and Bill Gross Stem Cell Research Center, University of California–Irvine, Irvine, California, USA;2. Department of Immunology, School of Medicine, Kyungpook National University, Daegu, Korea;1. Laboratoire des Substituts Cutanés, Hôpital Edouard Herriot, 5 place d’Arsonval, Pavillon i, 69437, Lyon, France;2. Laboratoire de Biologie Tissulaire et Ingénierie Thérapeutique, Institut de Biologie et Chimie des Protéines, UMR 5305, CNRS, Univ. Lyon 1, SFR BioSciences Gerland-Lyon Sud, 7 passage du Vercors, 69367, Lyon, France;3. LabSkin Creations, Hôpital Edouard Herriot, 5 place d’Arsonval, Pavillon i, 69437, Lyon, France |
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Abstract: | Background aimsThe aim of the present study was to evaluate the effects of air-liquid interface on the differentiation potential of human amnion epithelial cells (HAECs) to skin-like substitute in organotypic culture.MethodsHAECs at passage 1–2 were seeded onto a fibrin layer populated with human amnion mesenchymal cells to form the organotypic cultures. The organotypic HAECs were then cultured for 7, 14 and 21 d in two types of culture system: the submerged culture and the air-liquid interface culture. Cell morphogenesis was examined under the light and electron microscopes (transmission and scanning) and analyzed by immunohistochemistry.ResultsOrganotypic HAECs formed a single layer epithelium after 3 wk in submerged as well as air-liquid interface cultures. Ultrastructurally, desmosomes were observed in organotypic HAECs cultured in the air-liquid interface but not in the submerged culture. The presence of desmosomes marked the onset of early epidermal differentiation. Organotypic HAECs were positive against anti-CK18 and anti-CK14 in both the submerged and the air-liquid interface cultures. The co-expression of CK14 and CK18 suggested that differentiation of HAECs into skin may follow the process of embryonic skin development. However, weak expression of CK14 was observed after 2 and 3 wk of culture in air-liquid interface. CK10, involucrin, type IV collagen and laminin-5 expression was absent in organotypic HAECs. This observation reflects the initial process of embryonic epidermal differentiation and stratification.ConclusionsResults from the present study suggest that the air-liquid interface could stimulate early differentiation of organotypic HAECs to epidermal cells, with a potential use for skin regeneration. |
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