title: Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration.
abstract: OBJECTIVE
  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.
  
  
  RESULTS
  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.
authors.affiliation: Department of Pathology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur, Malaysia.
authors.forename: Simat Siti
authors.initials: SS
authors.lastname: Fatimah
authors.forename: Kienhui
authors.initials: K
authors.lastname: Chua
authors.forename: Geok Chin
authors.initials: GC
authors.lastname: Tan
authors.forename: Tengku Ibrahim
authors.initials: TI
authors.lastname: Azmi
authors.forename: Ay Eeng
authors.initials: AE
authors.lastname: Tan
authors.forename: Hayati
authors.initials: H
authors.lastname: Abdul Rahman
mesh_terms: D000650:Amnion / Q000166:cytology*; D015815:Cell Adhesion Molecules / Q000096:biosynthesis; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000096:biosynthesis; D003896:Desmosomes / Q000378:metabolism; D000078404:Epidermal Cells; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism; D009924:Organ Culture Techniques; D011498:Protein Precursors / Q000096:biosynthesis; D012038:Regeneration / Q000502:physiology*; D012879:Skin Physiological Phenomena; D014161:Transduction, Genetic; D014945:Wound Healing; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D024141:Collagen Type IV; D011498:Protein Precursors; D000097794:Kalinin; C032253:involucrin
keywords: air-liquid interface; epithelial stem cells; fibrin; human amnion–derived stem cells; organotypic culture; skin regeneration