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Xiaoqing Q Guo, Audrey E. K. Hutcheon, James D Zieske; Expression of TSP1 in Human Corneal Cells. Invest. Ophthalmol. Vis. Sci. 2018;59(9):1369.
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In previous studies, we found that transforming growth factor beta (TGFβ) was involved in corneal wound healing, and TGF-β1 stimulated the upregulation of thrombospondin 1 (TSP1) in human corneal epithelial and fibroblast cells (HCE and HCF, respectively). TSP1 has also been shown to localize in the Descemet’s membrane (DM) and endothelial cells, as well as being involved in corneal wound healing. In addition, wound healing in TSP1-/- mice after a penetrating wound resulted in persistent edema and wound gaping with no sign of DM reformation. Therefore, we hypothesized that TSP1 in human corneal endothelial cells (HCEndo) is important for wound healing.
HCEndo were cultured in Chen’s Endothelial Medium and treated with 2ng/ml TGF-β1 for 0, 1, 2, 3, 4, and 22 hours. Cells were processed for Western Blot (WB) and the membrane was probed with anti-TSP1 and β-actin, a loading and internal control. Data was quantitated and graphed.
The WB results showed that unlike HCE and HCF, HCEndo had no response to being stimulated with TGF-β1. Little, if any, TSP1 protein was detected by WB.
Unlike HCE and HCF, HCEndo did not respond to TGF-β1 stimulation to upregulate TSP1; therefore, the TSP1 that is found to be involved in wound healing of the cornea, must be coming from the HCE, HCF, and/or aqueous humor to reform the DM.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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