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Eddie Wang, Dong Wang, Andrew Geng, Ankur Maniar, Xiaohua Gong; A 3D Lens Cell Culture Platform to Mimic the Developing Lens. Invest. Ophthalmol. Vis. Sci. 2016;57(12):3060.
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© ARVO (1962-2015); The Authors (2016-present)
Currently, mechanistic study of lens biology relies on the use of animal models. Therefore, testing the roles and effects of small molecules, growth factors, and genetic alterations is low throughput and cost intensive. Our goal is to create a system for highly parallel and efficient in vitro synthesis of lens-like organoids that could greatly accelerate basic and applied lens research while decreasing the requirement for lab animals. The lens-like organoids will also open the door to real-time monitoring of molecular and cellular interactions.
We expanded primary tdTomato-H2B-GFP mouse lens epithelial cells (LECs) then embedded them in sacrifical hydrogel microspheres of ~200 microns in diameter. We embedded the spheres in hydrogels then dissolved the microspheres. The remaining cells were allowed to grow within the hollow spherical voids and exposed to basic fibroblast growth factor to induce differentiation. The LECs were imaged during and after culture by confocal microscopy.
LECs survived and proliferated after encapsulation and culture within the voids of hydrogels. Cell proliferation and morphology depended on the hydrogel stiffness and chemical properties and on the presence or absence of small molecule pathway inhibitors. Different stages of differentiated and mature fibers wre observed in the cultures.
This system allows for the synthesis of thousands of lens vesicle-like hollow spheres from a single mouse lens. These lens-like organoids recapitulate some of distinct features of fiber cell differentiation and maturation. Further optimization of growth factor treatments will allow for studying the dynamics of LEC differentiation and fiber cell elongation and maturation.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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