Journal
SCIENTIFIC REPORTS
Volume 8, Issue -, Pages -Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41598-018-22821-9
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Funding
- Natural Sciences and Engineering Research Council of Canada (NSERC) [418598]
- FRQ-S
- Fondation Antoine Turmel
- Fonds Wilbrod-Bherer de l'Universite Laval
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Human corneal endothelial cells (HCECs) easily become fibroblastic-like when cultured, rendering them unsuitable for tissue engineering of the cornea. Transforming growth factor beta (TGF-beta) could be a key factor in this phenomenon; however, TGF-beta is also known to maintain the endothelium in a quiescent state in vivo. This work aimed to compare the effects of TGF-beta 1 on the phenotype of HCECs during the proliferation and maturation phases. Our results show that addition of TGF-beta 1 during the active proliferation phase produced fibroblastic HCECs and loss of the cell junction markers ZO-1 and n-cadherin, independent from the presence of epidermal growth factor (EGF). By contrast, addition of TGF-beta 1 in maturation media containing few mitogens led to an endothelial phenotype and functional cell junctions as HCECs developed a high trans-endothelial resistance. Furthermore, addition of AG-1478, an epithelial growth factor receptor inhibitor, enhanced the gain of the endothelial phenotype and cell barrier function. Overall, these results show that TGF-beta 1 can be used to promote the formation of a typical leaky endothelial barrier during the maturation phase of cultured HCECs. A two-phase culture of HCECs using distinct proliferation and maturation media could also be key for developing ideal HCEC culture conditions.
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