In vitro generation of functional lens-like structures with relevance to age-related nuclear cataract.
OBJECTIVE
To investigate the capacity of lens epithelial cells, maintained in a modified explant culture system, to mimic normal patterns of lens cell differentiation and to regenerate lens structure and function.


METHODS
Lens epithelial explants were set up in pairs with their apical surfaces facing each other. These explant pairs (EPs) were then cultured in vitreous for up to 43 days to promote their growth and differentiation. Immunohistochemistry and conventional light and electron microscopy were used to assess structural and functional properties of the lens-like structures that developed from EPs.


RESULTS
EPs that were asymmetrically exposed to vitreous routinely produced biconvex, lens-like structures composed of ordered epithelial and fiber cells that were transparent and had some focusing and magnifying ability. In addition, characteristic of the lens in vivo, fiberlike cells that were peripherally situated in EPs contained markers of the relatively early stages of fiber differentiation, whereas centrally situated cells contained markers of terminally differentiated fibers. During long-term culture of the EPs, a central opacity appeared that had structural features similar to those reported for the early stages of human, age-related nuclear cataract.


CONCLUSIONS
This study shows that, given appropriate culture conditions, lens epithelial cells can regenerate ordered lens-like structures with functional properties. This system represents a valuable new tool for the investigation of factors involved in the generation of normal lens structure and function and lens opacification.
O'Connor
Michael D
MD
Save Sight Institute, University of Sydney, Sydney, New South Wales, Australia.
McAvoy
John W
JW
D000375:Aging / Q000502:physiology*; D000818:Animals; D001082:Aqueous Humor / Q000502:physiology; D002386:Cataract / Q000473:pathology*; D002417:Cattle; D002454:Cell Differentiation; D002478:Cells, Cultured; D003459:Crystallins / Q000378:metabolism; D004847:Epithelial Cells / Q000502:physiology* / Q000648:ultrastructure; D007908:Lens, Crystalline / Q000166:cytology / Q000502:physiology*; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D051381:Rats; D017208:Rats, Wistar; D012038:Regeneration / Q000502:physiology*; D014822:Vitreous Body / Q000502:physiology
D003459:Crystallins