Human lens epithelial cell proliferation in a protein-free medium.
OBJECTIVE
The ocular humors are relatively low in protein, yet cell growth in the human capsular bag still occurs after extracapsular cataract extraction (ECCE) surgery. This resilient growth gives rise to posterior capsule opacification (PCO) in a significant proportion (30%) of patients. This study compared the ability of human lens cells to proliferate in serum-supplemented and protein-free medium.


METHODS
Sham cataract operations were performed on human donor eyes. The capsular bag was dissected free, pinned flat on a petri dish, and incubated in Eagle's minimal essential medium (EMEM) alone or in EMEM supplemented with 10% fetal calf serum. Observations were made by phase-contrast microscopy. At the endpoint, capsules were studied by fluorescence or electron microscopy. Mitotic activity was identified using Bromo-2-deoxyuridine labeling and detection techniques. When required, an intraocular lens was implanted when surgery was performed.


RESULTS
It was found that human lens cells from a wide age spectrum of donors proliferate and migrate on the lens capsule in the absence of added protein. The rate of growth was age-dependent, such that the posterior capsule was completely confluent after 8.0 +/- 0 days (n = 3) and 24.4 +/- 5.3 days (n = 3) for donor lenses aged < 40 years and > 60 years, respectively. The outgrowth of epithelial cells gave rise to capsular contraction, wrinkling, and increased light scatter. Growth on the anterior surface of the intraocular lens was less prolific than on the posterior capsule.


CONCLUSIONS
The protein-free model replicates many features of clinically-observed PCO. The resilient cell growth on the natural collagen capsule explains the high prevalence of PCO, especially in younger patients, and suggests that inflammation and external growth factors are not necessary for PCO. Furthermore, the protein-free capsular bag system can be used to explore fundamental questions concerning the autocrine control of lens epithelial cell survival and growth.
Wormstone
I M
IM
School of Biological Sciences, University of East Auglia, Norwich, UK.
Liu
C S
CS
Rakic
J M
JM
Marcantonio
J M
JM
Vrensen
G F
GF
Duncan
G
G
D000328:Adult; D001973:Bromodeoxyuridine; D002455:Cell Division; D016895:Culture Media, Serum-Free; D004247:DNA / Q000096:biosynthesis; D004261:DNA Replication; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007903:Lens Capsule, Crystalline / Q000737:chemistry / Q000166:cytology; D007908:Lens, Crystalline / Q000737:chemistry / Q000166:cytology*; D007910:Lenses, Intraocular; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D014746:Vimentin / Q000032:analysis
D016895:Culture Media, Serum-Free; D014746:Vimentin; D004247:DNA; D001973:Bromodeoxyuridine