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KMID : 1377020130100060362
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Tissue Engineering and Regenerative Medicine
2013 Volume.10 No. 6 p.362 ~ p.370
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Microarray analysis of oral mucosal epithelial cell sheet
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Fawzia Bardag-Gorce
Joan Oliva
Andrew Wood
Hope Niihara
Andrew Makalinao
Sean Sabino
Derek Pan
Jacquelyn Thropay
Hiroyuki Sota
Yutaka Niihara
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Abstract
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Corneal epithelium regeneration using autologous oral mucosal epithelial cell sheet is a successful new approach in corneal therapy. In the present study, gene expression profiling was performed to characterize engineered cell sheets. Cell sheets were obtained by culturing isolated rabbit oral mucosal epithelial cells on a thermoresponsive cultureware (UpCell®, CellSeed Inc. Japan). H&E staining of cell sheets showed a multistratified epithelium, similar to corneal epithelium. DeltaN-p63 stained positive in the basal cells, indicating that cell sheets have renewal capacity. Microarray analysis of these cell sheets showed that only 160 genes out of 43,000 rabbit probes listed on the microarray chip were identified. We first identified the extracellular matrix group of genes and found that matrix metalloproteinase MMP-1, MMP-3 and MMP-12, known to promote angiogenesis, were down regulated, while MMP-13 and collagen type VIII alpha 1 (COL8A1), proteins involved in wound healing, were up regulated. Tissue inhibitors of metalloproteinase TIMP-1 and TIMP-3, anti-angiogenic factors, were also identified. Gap junction protein A7 (GJA7 or Connexin 45) was found up regulated, indicating that cell sheets have developed well preserved cell-cell interactions. Alcohol dehydrogenase 5 (ADH class III) and aldehyde dehydrogenase (ALDH1A1), involved in protecting the cornea against oxidative stress induced by UV radiation, were also found up regulated. In conclusion, microarray analysis has led us to identify new target molecules and their subsequent biochemical analysis indicated how the composite cell sheets are advantageous to the original isolated cells in terms of the integrity and potency of corneal epithelial grafts without any scaffolds.
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KEYWORD
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oral, mucosal, epithelium, cell sheet, engineering, cornea
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