Value of human amniotic epithelial cells in tissue engineering for cornea

Simat Siti Fatimah, Sook Luan Ng, Chua Kien Hui, Abdul Rahman Hayati, Ay Eeng Tan, Geok Chin Tan

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Human amniotic epithelial cells (hAECs) are potentially one of the key players in tissue engineering due to their easy availability. The aim of the present study was to develop an optimal isolation and transportation technique, as well as to determine the immunophenotype and epithelial gene expression of hAECs. Amnion was mechanically peeled off from the chorion and digested with trypsin-ethylenediaminetetraacetic acid. The isolated hAECs were cultured in medium containing 10 ng/mL epidermal growth factor until P4. The epithelial gene expression, cell surface antigen and protein expression of hAECs were analyzed by quantitative polymerase chain reaction, flow cytometry and immunocytochemistry. hAECs were also cultured in adipogenic, osteogenic and neurogenic induction media. The best cell yield of hAECs was seen in the digestion of 15 pieces of amnion (2 × 2 cm) and isolated 30 min after digestion with trypsin. F12:Dulbecco's modified eagle medium was the best medium for short term storage at 4 °C. hAECs expressed CD9, CD44, CD73 and CD90, and negligibly expressed CD31, CD34, CD45 and CD117. After serial passage, CK3, CK19 and involucrin gene expressions were upregulated, while p63, CK1 and CK14 gene expressions were downregulated. Sustained gene expressions of integrin β1 and CK18 were observed. At initial culture, these cells might have stem-like properties. However, they differentiated after serial passage. Nonetheless, hAECs have epithelial stem cell characteristics and have the potential to differentiate into corneal epithelial cells. Further investigations are still needed to elucidate the mechanism of differentiation involved and to optimize the culture condition for long term in vitro culture.

Original languageEnglish
Pages (from-to)141-151
Number of pages11
JournalHuman Cell
Volume23
Issue number4
DOIs
Publication statusPublished - Nov 2010
Externally publishedYes

Fingerprint

Tissue Engineering
Cornea
Epithelial Cells
Gene Expression
Serial Passage
Amnion
Trypsin
Digestion
Chorion
Eagles
Surface Antigens
Epidermal Growth Factor
Integrins
Edetic Acid
Flow Cytometry
Membrane Proteins
Stem Cells
Down-Regulation
Immunohistochemistry
Polymerase Chain Reaction

Keywords

  • Amniotic epithelium
  • Cornea
  • Placenta
  • Regenerative medicine
  • Stem cells

ASJC Scopus subject areas

  • Cell Biology
  • Cancer Research

Cite this

Value of human amniotic epithelial cells in tissue engineering for cornea. / Fatimah, Simat Siti; Ng, Sook Luan; Kien Hui, Chua; Hayati, Abdul Rahman; Tan, Ay Eeng; Tan, Geok Chin.

In: Human Cell, Vol. 23, No. 4, 11.2010, p. 141-151.

Research output: Contribution to journalArticle

Fatimah, Simat Siti ; Ng, Sook Luan ; Kien Hui, Chua ; Hayati, Abdul Rahman ; Tan, Ay Eeng ; Tan, Geok Chin. / Value of human amniotic epithelial cells in tissue engineering for cornea. In: Human Cell. 2010 ; Vol. 23, No. 4. pp. 141-151.
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