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H. Nanba, Y. Kashiwagi, H. Takamura, T. Yamamoto, H. Yamashita; Effects of Platelet Derived Growth Factor-BB (PDGF-BB) on Gene Expression Patterns in Human Scleral Stromal Cells and Corneal Stromal Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4443. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate gene expression patterns after PDGF-BB stimulation in scleral stromal cells and to compare the difference among the above cells and corneal stromal cells and human epidermal fibroblast cells.
Human scleral stromal cells and corneal stromal cells were cultured from the explants of human materials, and were transformed by human papilloma virus 16 (HPV16) bector. To compare the characteristics, human epidermal fibroblasts were purchased (NB1RGB, obtained from Riken Cell Bank). Changes of gene expression patterns after PDGF-BB stimulation (10 ng/ml, 24 hours) were analyzed by the DNA microarray (Human Genome U113 Plus 2.0 Array, Affymetrix) in scleral stromal cells. To confirm the expression changes of the selected genes, reverse transcription-polymerase chain reaction (RT-PCR) was performed in the above mentioned 3 types of cells.
The expression levels of thrombomodulin (TM), hepatocyte growth factor (HGF) and matrix metalloproteinase 16 (MMP16) genes were increased after the results of DNA microarray in the scleral cells, and the increase was confirmed by RT-PCR. At the baseline status, the expression of TM gene was lower in the epidermal fibroblasts than the others, that of HGF gene was higher in the scleral cell, and that of MMP16 gene was lower in the corneal cell. The expressions of TM gene and MMP16 increased in all the cell types. HGF expression level increased in the scleral cells and the corneal cells, not in the epidermal fibroblasts. Although expression levels of tissue factor pathway inhibitor 2, versican, E2 transcription factor 8, Apolipoprotein L6 and epiphycan were also increased in the DNA microarray, there were no significant differences among them.
It is shown that the gene expression patterns after PDGF-BB stimulation were different among in scleral stromal cells, corneal stromal cells and epidermal fibroblast cells.
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