Purchase this article with an account.
Bharat Kumar, Matthew Aaron Reilly; Maintaining Epithelial Cell Viability in Whole Lens Cultures ex vivo. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3639.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Lens explants have previously been used to study the behavior of lens epithelial cell (LEC) in vitro but show altered LEC morphology. We therefore evaluated the viability of LECs when culturing whole lenses in vitro in the presence and absence of vitreous humor.
Pairs of freshly enucleated porcine eyes were dissected and the crystalline lens was removed. One lens from each pair was cultured in growth media for 24 hours while the other was immediately dissected. Lenses were cultured in M199 media which, in some cases, was augmented with vitreous humour in a 1:4 ratio. LEC viability was quantified using a hemocytometer after removing the fiber cell bundle and trypsinizing the anterior lens capsule. The viable cell counts from the cultured lenses were normalized to data from the fresh lenses of each pair.
The normalized viable cell population for lenses cultured in the vitreous enhancedmedia was 1.07. The normalized population for the control media was 0.87. Preliminary results indicate that the vitreous-enhanced media played a role in maintaining lens epithelial cell viability (Figure 1).
Viable cell populations in lenses cultured in enhanced media remained consistent after the culture period in contrast to the rapid decrease in lenses cultured in the control media. This information will enable longer-term cultures to determine the influence of additional factors in LEC viability and proliferation.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
Figure 1. Normalized viable cell populations in lenses cultured in enhanced (left) and control (right) media relative to the viable cell population in fresh lenses
This PDF is available to Subscribers Only