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RISS 인기검색어
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- 저자 : Hiroyuki Sota (검색결과 2 건)
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Joan Oliva,Fawzia Bardag-Gorce,Andrew Wood,Hiroyuki Sota,Yutaka Niihara 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.5
Autologous stem cell transplantation for eye diseases is immunologically preferable to avoid allograft rejection. However, the fate of the grafted cells has never been studied. Here, we propose to use 19F-perfluorocarbon magnetic resonance imaging tracer agent, to label cell sheet in vitro. This labeling enables non-invasive visualization of possible migration of grafted cells. Oral mucosal epithelial cells were isolated from rabbit oral mucosal epithelium and were cultivated in a thermo-responsive surface to engineer a multilayer cell sheet. Different concentrations of 19F-perfluorocarbon were added to the cell sheet culture media, one or two times. Cells were analyzed in a 7 T nuclear magnetic resonance to determine the labeling efficiency. We found that 10 mg/mL and two incubations with 19F-perfluorocarbon were the optimal condition for labeling. H&E and immunocytochemistry showed that labeling did not affect the expression of cell sheets specific markers (CK4, CK13, connexin43, E-cadherin). Furthermore, no significant effects were observed on the number of cells and the cell viability, making 19F-perfluorocarbon suitable for cell tracking, with no side effects.
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Microarray Analysis of Oral Mucosal Epithelial Cell Sheet
Fawzia Bardag-Gorce,Joan Oliva,Hope Niihara,Andrew Makalinao,Sean Sabino,Derek Pan,Jacquelyn Thropay,Hiroyuki Sota,Yutaka Niihara,Andrew Wood 한국조직공학과 재생의학회 2013 조직공학과 재생의학 Vol.10 No.6
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 engi-neered cell sheets. Cell sheets were obtained by culturing isolated rabbit oral mucosal epithelial cells on a thermo-responsive cultureware (UpCell®, CellSeed Inc. Japan). H&E staining of cell sheets showed a multistratified epi-thelium, 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 regu-lated, while MMP-13 and collagen type VIII alpha 1 (COL8A1), proteins involved in wound healing, were up reg-ulated. 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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