Ex vivo canine lens capsular sac explants

Graefes Arch Clin Exp Ophthalmol. 2000 Aug;238(8):708-14. doi: 10.1007/s004170000158.

Abstract

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.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / metabolism
  • Animals
  • Biomarkers
  • Cataract / metabolism
  • Cataract / pathology
  • Cell Division
  • Cells, Cultured
  • Dogs
  • Enzyme-Linked Immunosorbent Assay
  • Insulin-Like Growth Factor I / metabolism*
  • Lens Capsule, Crystalline / cytology
  • Lens Capsule, Crystalline / growth & development
  • Lens Capsule, Crystalline / metabolism*
  • Transferrin / metabolism*
  • Transforming Growth Factor beta / metabolism*
  • Transforming Growth Factor beta2
  • Vimentin / metabolism

Substances

  • Actins
  • Biomarkers
  • TGFB2 protein, human
  • Transferrin
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta2
  • Vimentin
  • Insulin-Like Growth Factor I