BACKGROUND: Lens capsular sac explants from human cadaver eyes were used to investigate posterior capsular opacification (PCO). The purpose of this study was to characterize a similar model using canine tissue and to determine whether transferrin (Tf), transforming growth factor beta-2 (TGF-beta2), and insulin-like growth factor-1 (IGF-1) are secreted by lens epithelial cells (LEC) of these ex vivo sacs. METHODS: The lens from canine eyes was removed by extracapsular cataract extraction, the lens sac dissected free, pinned to a petri dish, and cultured in either serum-supplemented or serum-free medium. Morphologic characteristics and growth rate to confluence on the posterior capsule were studied by phase-contrast microscopy. Vimentin, alpha smooth muscle actin, and panTGF-beta expression by LEC were determined by immunohistochemistry. Tf, TGF-beta2, and IGF-1 levels were measured by ELISA in the supernatant of sacs cultured in serum-free medium. RESULTS: The mean time to confluence of LEC onto the posterior capsule was 5.4+/-1.1 days (n=22) and 14.7+/-3.7 days (n=14) for sacs in serum-supplemented and serum-free medium, respectively. Following development of confluence, explants displayed opacification and light scatter from cellular proliferation and capsular contraction. Confluent LEC expressed vimentin, alpha smooth muscle actin, and TGF-beta2, and both Tf and TGF-beta2 were secreted into the culture supernatant. CONCLUSION: Canine lens sac explants have characteristics virtually identical to those of human origin, and appear to be a useful alternative tissue source for this model when human cadaver eyes are unavailable. Tf and TGFbeta-2, but not IGF-1, are secreted by LEC in explanted lens sacs and may influence the proliferation and metaplasia of LEC during the development of PCO.
BACKGROUND: Lens capsular sac explants from human cadaver eyes were used to investigate posterior capsular opacification (PCO). The purpose of this study was to characterize a similar model using canine tissue and to determine whether transferrin (Tf), transforming growth factor beta-2 (TGF-beta2), and insulin-like growth factor-1 (IGF-1) are secreted by lens epithelial cells (LEC) of these ex vivo sacs. METHODS: The lens from canine eyes was removed by extracapsular cataract extraction, the lens sac dissected free, pinned to a petri dish, and cultured in either serum-supplemented or serum-free medium. Morphologic characteristics and growth rate to confluence on the posterior capsule were studied by phase-contrast microscopy. Vimentin, alpha smooth muscle actin, and panTGF-beta expression by LEC were determined by immunohistochemistry. Tf, TGF-beta2, and IGF-1 levels were measured by ELISA in the supernatant of sacs cultured in serum-free medium. RESULTS: The mean time to confluence of LEC onto the posterior capsule was 5.4+/-1.1 days (n=22) and 14.7+/-3.7 days (n=14) for sacs in serum-supplemented and serum-free medium, respectively. Following development of confluence, explants displayed opacification and light scatter from cellular proliferation and capsular contraction. Confluent LEC expressed vimentin, alpha smooth muscle actin, and TGF-beta2, and both Tf and TGF-beta2 were secreted into the culture supernatant. CONCLUSION:Canine lens sac explants have characteristics virtually identical to those of human origin, and appear to be a useful alternative tissue source for this model when human cadaver eyes are unavailable. Tf and TGFbeta-2, but not IGF-1, are secreted by LEC in explanted lens sacs and may influence the proliferation and metaplasia of LEC during the development of PCO.
Authors: Jennifer L Davis; Na Young Yi; Jacklyn H Salmon; Anna N Charlton; Carmen M H Colitz; Brian C Gilger Journal: J Ocul Pharmacol Ther Date: 2012-02-28 Impact factor: 2.671
Authors: Heather L Chandler; Terah R Webb; Curtis A Barden; Mirunalni Thangavelu; Samuel K Kulp; Ching-Shih Chen; Carmen M H Colitz Journal: Mol Vis Date: 2010-10-29 Impact factor: 2.367