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

Simat Siti Fatimah, Chua Kien Hui, Geok Chin Tan, Tengku Ibrahim Azmi, Ay Eeng Tan, Hayati Abdul Rahman

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

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 1e2 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 airliquid 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.

Original languageEnglish
Pages (from-to)1030-1041
Number of pages12
JournalCytotherapy
Volume15
Issue number8
DOIs
Publication statusPublished - 2013

Fingerprint

Artificial Skin
Amnion
Regeneration
Air
Epithelial Cells
Desmosomes
Skin
Collagen Type IV
Fibrin
Morphogenesis
Embryonic Development
Epithelium
Immunohistochemistry

Keywords

  • Air-liquid interface
  • Epithelial stem cells
  • Fibrin
  • Human amnionederived stem cells
  • Organotypic culture
  • Skin regeneration

ASJC Scopus subject areas

  • Cell Biology
  • Cancer Research
  • Transplantation
  • Genetics(clinical)
  • Oncology
  • Immunology and Allergy
  • Immunology

Cite this

Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration. / Fatimah, Simat Siti; Kien Hui, Chua; Tan, Geok Chin; Azmi, Tengku Ibrahim; Tan, Ay Eeng; Rahman, Hayati Abdul.

In: Cytotherapy, Vol. 15, No. 8, 2013, p. 1030-1041.

Research output: Contribution to journalArticle

Fatimah, Simat Siti ; Kien Hui, Chua ; Tan, Geok Chin ; Azmi, Tengku Ibrahim ; Tan, Ay Eeng ; Rahman, Hayati Abdul. / Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration. In: Cytotherapy. 2013 ; Vol. 15, No. 8. pp. 1030-1041.
@article{7792966f36e74401ac990dee5f734dfe,
title = "Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration",
abstract = "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 1e2 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 airliquid 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.",
keywords = "Air-liquid interface, Epithelial stem cells, Fibrin, Human amnionederived stem cells, Organotypic culture, Skin regeneration",
author = "Fatimah, {Simat Siti} and {Kien Hui}, Chua and Tan, {Geok Chin} and Azmi, {Tengku Ibrahim} and Tan, {Ay Eeng} and Rahman, {Hayati Abdul}",
year = "2013",
doi = "10.1016/j.jcyt.2013.05.003",
language = "English",
volume = "15",
pages = "1030--1041",
journal = "Cytotherapy",
issn = "1465-3249",
publisher = "Informa Healthcare",
number = "8",

}

TY - JOUR

T1 - Organotypic culture of human amnion cells in air-liquid interface as a potential substitute for skin regeneration

AU - Fatimah, Simat Siti

AU - Kien Hui, Chua

AU - Tan, Geok Chin

AU - Azmi, Tengku Ibrahim

AU - Tan, Ay Eeng

AU - Rahman, Hayati Abdul

PY - 2013

Y1 - 2013

N2 - 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 1e2 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 airliquid 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.

AB - 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 1e2 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 airliquid 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.

KW - Air-liquid interface

KW - Epithelial stem cells

KW - Fibrin

KW - Human amnionederived stem cells

KW - Organotypic culture

KW - Skin regeneration

UR - http://www.scopus.com/inward/record.url?scp=84890901867&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84890901867&partnerID=8YFLogxK

U2 - 10.1016/j.jcyt.2013.05.003

DO - 10.1016/j.jcyt.2013.05.003

M3 - Article

C2 - 23830235

AN - SCOPUS:84890901867

VL - 15

SP - 1030

EP - 1041

JO - Cytotherapy

JF - Cytotherapy

SN - 1465-3249

IS - 8

ER -