title: Rapamycin Prolongs the Survival of Corneal Epithelial Cells in Culture.
abstract: Rapamycin has previously been shown to have anti-aging effects in cells and organisms. These studies were undertaken to investigate the effects of rapamycin on primary human corneal epithelial cells in vitro. Cell growth and viability were evaluated by bright field microscopy. Cell proliferation and cycle were evaluated by flow cytometry. The expression of differentiation markers was evaluated by quantitative PCR and Western blot. Senescence was evaluated by senescence-associated β-Galactosidase staining and by Western blot analysis of p16. Apoptosis was evaluated by a TUNEL assay. The results demonstrated that primary HCEC treated with rapamycin had lower proliferation but considerably longer survival in vitro. Rapamycin-treated cells maintained a higher capacity to proliferate after removal of rapamycin and expressed more keratin 14, N-Cadherin, DeltaNp63 and ABCG2, and less keratin 12, consistent with their less differentiated state. Rapamycin treated cells demonstrated less senescence by X-β-Gal SA staining and by lower expression of p16. Apoptosis was also lower in the rapamycin treated cells. These results indicate that rapamycin treatment of HCEC prevents the loss of corneal epithelial stem/progenitor cells to replicative senescence and apoptosis. Rapamycin may be a useful additive for ex vivo expansion of corneal epithelial cells.
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Sanaz
authors.initials: S
authors.lastname: Gidfar
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Farnoud Y
authors.initials: FY
authors.lastname: Milani
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Behrad Y
authors.initials: BY
authors.lastname: Milani
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Xiang
authors.initials: X
authors.lastname: Shen
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Medi
authors.initials: M
authors.lastname: Eslani
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Ilham
authors.initials: I
authors.lastname: Putra
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Huvard
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Hossein
authors.initials: H
authors.lastname: Sagha
authors.affiliation: Department of Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago IL, USA.
authors.forename: Ali R
authors.initials: AR
authors.lastname: Djalilian
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D017209:Apoptosis / Q000187:drug effects; D015415:Biomarkers / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D019573:Epithelium, Corneal / Q000166:cytology*; D006801:Humans; D007249:Inflammation / Q000473:pathology; D016850:Limbus Corneae / Q000166:cytology; D008875:Middle Aged; D020123:Sirolimus / Q000494:pharmacology*; D055815:Young Adult
chemical_list: D015415:Biomarkers; D020123:Sirolimus