Purpose: To develop and evaluate a organ culture model for investigating the EMT of human lens capsules in vitro.
Methods. The cataract surgery, including anterior capsulorhexis, nucleus hydroexpression, and aspiration of lens fibers, was performed on donor eyes. The capsular bag was dissected free, pinned flat on a plastic culture dish, covered with DMEM/F12 supple-mented with 10% fetal calf serum and observed by phase-contrast and dark-field microscopy for as long as 100 days. At the different time-point, capsules were examined by fluorescence microscopy, western blot and QPCR for SMA, vimentin, and E-cadherin.
Results: After 2 to 3 days, cells were normally observed growing from the rhexis onto the posterior capsule. Growth proceeded rapidly so that the posterior capsule, for example, was totally covered by a confluent monolayer of cells at 5-7 days for capsules. Capsular wrinkles became increasingly apparent as time progressed, and an increase in capsular tension also occurred. E-cadherin were positive on day0, negative on day7,day14, and positive again on day28. SMA were negative on day0, and positive on day7,day14 and day28. Vimentin were positive all the time-points.
Conclusions: The model will be useful to investigate the EMT strategies for posterior capsule opacification. |
