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Takafumi Miyatani, Hiroshi Tanaka, Takahiro Nakamura, Tomoko Horikiri, Megumi Kishida, YOICHIRO SHINKAI, Kentaro Kojima, Chie Sotozono; Morphological change and migration activity of human Müller cells on the amniotic membrane. Invest. Ophthalmol. Vis. Sci. 2021;62(8):3635.
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© ARVO (1962-2015); The Authors (2016-present)
It has recently been reported that amniotic membrane (AM) transplantation is effective for macular hole (MH) closure and improvement of visual function in refractory MH cases. In this study, we investigated the effects of AM on the morphology and migration of cultivated Müller cells (MIO-M1 cells, an immortalized model of Müller glial cells, University College London, London, UK).
MIO-M1 were cultured using Dulbecco's modified eagle medium with 10% fetal bovine serum in a humidified 5% CO2 environment at 37°C. AM was then placed in 24-well plates to create an epithelial-side-up group (epithelial group), a chorion-side-up group (chorion group), and a control group on glass slides (n = 10 each). Then, silicon (3mm in diameter) was placed in the center of each. Each group was then seeded with 0.5 x 105 MIO-M1 cells, with the silicon being removed after 24 hours. Next, phase-contrast microscopy images were taken immediately and at 72-hours post removal of the silicon, and actin filaments were visualized with phalloidin staining. Via examination of the obtained images, the cell morphology was analyzed and the migration ability of the cells was evaluated and compared between the three groups by measuring the cell migration area using ImageJ software.
The MIO-M1 cells showed a bipolar morphology with pseudopodia in the epithelial group and control group, and a nearly circular morphology in the chorionic villus group. In the control group, epithelial group, and chorionic group, the cell migration area was 0.34±0.26 mm2, 1.14±0.48 mm2, and 0.04±0.03 mm2 (mean±SD), respectively, with the ability of cell migration being significantly higher in the epithelial group (P<0.05).
Human Müller cells on the chorionic side showed a different morphology from the control and epithelial side, and their migration ability was lower than the epithelial-side cells. When performing AM transplantation for the treatment of MH, positioning the epithelial side of the AM toward the vitreous cavity may promote the migration of Müller cells and assist in the closure of the MH.
This is a 2021 ARVO Annual Meeting abstract.
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