Abstract
ABSTRACTCataracts cause vision loss and blindness by impairing the ability of the ocular lens to focus light onto the retina. Various cataract risk factors have been identified, including drug treatments, age, smoking and diabetes. However, the molecular events responsible for these different forms of cataract are ill-defined, and the advent of modern cataract surgery in the 1960s virtually eliminated access to human lenses for research. Here, we demonstrate large-scale production of light-focusing human micro-lenses from spheroidal masses of human lens epithelial cells purified from differentiating pluripotent stem cells. The purified lens cells and micro-lenses display similar morphology, cellular arrangement, mRNA expression and protein expression to human lens cells and lenses. Exposing the micro-lenses to the emergent cystic fibrosis drug Vx-770 reduces micro-lens transparency and focusing ability. These human micro-lenses provide a powerful and large-scale platform for defining molecular disease mechanisms caused by cataract risk factors, for anti-cataract drug screening and for clinically relevant toxicity assays.
Highlights
During embryogenesis, the ocular lens arises from the lens placode in the surface ectoderm opposite the optic cup (Mann, 1964; Tholozan and Quinlan, 2007)
The exact process can differ between vertebrate species, key lens features shared by vertebrates include an anterior lens epithelial cell (LEC) monolayer expressing α-crystallins overlying a mass of lens fibre cells expressing α, βand γ-crystallins (Thomson and Augusteyn, 1985)
To improve the suitability of in vitro lens regeneration for targeted and large-scale cataract studies, we investigated human pluripotent stem cells as a source of LECs
Summary
The ocular lens arises from the lens placode in the surface ectoderm opposite the optic cup (Mann, 1964; Tholozan and Quinlan, 2007). Invagination of the lens placode is followed by formation of the lens vesicle – a spherical LEC monolayer surrounding an acellular Differentiation of the posterior LECs into lens fibre cells fills the lens vesicle lumen to establish the basic lens architecture For decades these features have provided a framework for in vitro lens and cataract studies using explanted primary rat LECs. For example, our group reported in vitro regeneration of light-focusing rat lenses from paired rat LEC monolayers arranged to mimic lens vesicles (O’Connor and McAvoy, 2007). Continued culture of these regenerated rat lenses resulted in formation of a human-like cataract, as seen by reduced light transmission and reduced focusing ability
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