Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration

Siti Fatimah Simat and Chua, Kien hui and Geok, Chin Tan and Tengku Ibrahim Azmi and Hayati Abdul Rahman (2013) Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration. Cytotherapy, 15. pp. 1030-1041. ISSN 1465-3249


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Background aims The 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. Methods HAECs 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. Result Organotypic 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. Conclusions Results 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.

Item Type: Article
Keyword: human fibrin as a biomaterial, Organotypic culture of human amnion-derived cells, skin keratinocytes and dermal fibroblasts
Subjects: Q Science > QM Human anatomy
Date Deposited: 29 Jan 2014 11:27
Last Modified: 11 Oct 2017 14:56

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