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Graefe's Archive for Clinical and Experimental Ophthalmology

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01-09-2018 | Retinal Disorders

Premacular membranes in tissue culture

Authors: Denise Vogt, Franziska Vielmuth, Christian Wertheimer, Felix Hagenau, Stefanie R. Guenther, Armin Wolf, Volker Spindler, Siegfried G. Priglinger, Ricarda G. Schumann

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 9/2018

Abstract

Purpose

To investigate integrity and characteristics of human premacular membranes (PMM) with and without standard tissue culturing using mechanical traction.

Methods

Premacular membranes were harvested from 32 eyes of 32 patients with idiopathic macular pucker during standard vitrectomy. By flat-mount preparation with phase contrast and interference microscopy, specimens were prepared for time-lapse microscopy, immunocytochemistry, and transmission electron microscopy. Sixteen of 32 specimens were held in tissue culture with tangential traction by using entomological pins. Of these, specimens of 7 eyes were analyzed with and without tissue culturing for comparison. Primary antibodies were used for myofibroblasts, hyalocytes, macro-/microglial cells, and retinal pigment epithelial and immune cells.

Results

Hyalocytes, macroglia, and microglia composed the main cell composition of surgically removed PMM. Correlation of time-lapse microscopy with immunofluorescence microscopy identified fast and unidirectional moving small round cells as microglia. Slowly moving elongated large cells were characterized as alpha-smooth muscle actin (α-SMA)-positive myofibroblasts. Following tissue culturing with tangential stretch, enhanced positive immunolabelling for α-SMA and integrins-αv was seen. All other labelling results were demonstrated to be similar with pre-culture conditions. Ultrastructural analysis revealed fibroblasts, myofibroblasts, and proliferation of glial cells following tissue culture.

Conclusion

This study demonstrates abundance of fibroblasts, myofibroblasts, and glial cells in PMM from idiopathic macular pucker following tissue culture with tangential stretch application. We found enhanced contractive properties of the cultured PPM that appear to indicate transdifferentiation of the cell composition. This in vitro model may improve understanding of pathogenesis in traction maculopathies and help to establish further anti-fibrosis treatment strategies.

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Title: Premacular membranes in tissue culture
Authors: Denise Vogt
Franziska Vielmuth
Christian Wertheimer
Felix Hagenau
Stefanie R. Guenther
Armin Wolf
Volker Spindler
Siegfried G. Priglinger
Ricarda G. Schumann
Publication date: 01-09-2018
Publisher: Springer Berlin Heidelberg
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 9/2018
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-018-4033-6

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Springer Medicine

Premacular membranes in tissue culture

Abstract

Purpose

Methods

Results

Conclusion

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

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