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S. Lee, J.H. Oh, J.H. Yoon, S.J. Cho, J.H. Lee, S.H. Lee, M.I. Lee; Differentiation and Transdifferentiation of Lens Epithelial Cells In a Capsular Bag Model . Invest. Ophthalmol. Vis. Sci. 2004;45(13):369.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: In the capsular bag model where lens epithelial cells(LECs) are directly cultured on the lens capsule, we observed whether LECs would differentiate into lens fibers and transdifferentiate into fibroblasts. Methods:We cultured the capsular bags of porcine eyes with in–the–bag implantation of a PMMA tension ring. We observed whether LECs would differentiate into lens fibers or transdifferentiate into fibroblasts with a phase contrast microscope, a light microscope(LM) and an electron microscope(EM). In addition, the expressions of αA–crystallin, the marker expressed when LECs are differentiated into lens fibers and α–smooth muscle actin, when they are transdifferentiated into fibroblasts were examined using western blot. Results: According to the observation using a phase contrast microscope, the proliferation and migration of LECs were seen by 2–8 days after culture. Some LECs elongated gradually and their cytoplasm became transparent showing the differentiation into lens fibers by 7–25 days after culture. By 7–15 days after culture, some LECs transdifferentiated into fibroblasts with their morphology changing into spindle–shape. From LM and EM observations it would be appropriate to express view that changes of each cell were lens fiber, and fibroblast–like cells. According to western blot performed immediate after culture and 3, 6, 10, 15, 20, 25, and 30 after culture, the expression of αA–crystallin started immediately after culture, was the highest by 10 days after culture, and maintained until 20 days after culture when it gradually disappeared. The expression of α–smooth muscle actin was started by 3 days after culture, the highest by 10 days after culture with its high expression maintained until 30 days after culture. Conclusions: LECs culture using the capsular bag model would be effective in researching the process of LEC differentiation and transdifferentiation, the prevention of cataract, and the pathophysiologic mechanism and inhibition of after–cataract.
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