Feet on the ground: Physical support of the inner retina is a strong determinant for cell survival and structural preservation in vitro.
(2014) In Investigative Ophthalmology & Visual Science 55(4). p.2200-2213- Abstract
- Purpose: The purpose of this study was to explore the importance of local physical tissue support for homeostasis in the isolated retina. Methods: Full-thickness retinal sheets were isolated from adult porcine eyes. Retinas were cultured for 5 or 10 days using the previously established explant protocol with photoreceptors positioned against the culture membrane (porous polycarbonate) or the Müller cell endfeet and inner limiting membrane (ILM) apposed against the membrane. The explants were analyzed morphologically using hematoxylin and eosin staining, immunohistochemistry, TUNEL labeling, and transmission electron microscopy (TEM). Results: Standard cultures displayed a progressive loss of retinal lamination and extensive cell death,... (More)
- Purpose: The purpose of this study was to explore the importance of local physical tissue support for homeostasis in the isolated retina. Methods: Full-thickness retinal sheets were isolated from adult porcine eyes. Retinas were cultured for 5 or 10 days using the previously established explant protocol with photoreceptors positioned against the culture membrane (porous polycarbonate) or the Müller cell endfeet and inner limiting membrane (ILM) apposed against the membrane. The explants were analyzed morphologically using hematoxylin and eosin staining, immunohistochemistry, TUNEL labeling, and transmission electron microscopy (TEM). Results: Standard cultures displayed a progressive loss of retinal lamination and extensive cell death, with activated, hypertrophic Müller cells. In contrast, explants cultured with the ILM facing the membrane displayed a maintenance of the retinal laminar architecture, and a statistically significant attenuation of photoreceptor and ganglion cell death. TEM revealed intact synapses as well as preservation of normal cellular membrane structures. Immunohistochemistry showed no signs of Müller cell activation (GFAP), with maintained expression of important metabolic markers (GS, bFGF). Conclusion: Providing physical support to the inner but not the outer retina appears to prevent the tissue collapse resulting from perturbation of the normal biomechanical milieu in the isolated retinal sheet. Using this novel paradigm, gliotic reactions are attenuated, and metabolic processes vital for tissue health are preserved which significantly increases neuronal cell survival. This finding opens up new avenues of adult retinal tissue culture research, and increases our understanding of pathological reactions in biomechanically related conditions in vivo. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4383858
- author
- Taylor, Linnéa LU ; Arnér, Karin LU ; Holmgren Taylor, Ingrid and Ghosh, Fredrik LU
- organization
- publishing date
- 2014
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Investigative Ophthalmology & Visual Science
- volume
- 55
- issue
- 4
- pages
- 2200 - 2213
- publisher
- Association for Research in Vision and Ophthalmology Inc.
- external identifiers
-
- pmid:24595389
- wos:000335913100025
- scopus:84898746584
- pmid:24595389
- ISSN
- 1552-5783
- DOI
- 10.1167/iovs.13-13535
- language
- English
- LU publication?
- yes
- id
- 9e2a5348-8239-4cf3-b7b9-cd18e15751ce (old id 4383858)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/24595389?dopt=Abstract
- date added to LUP
- 2016-04-01 09:53:21
- date last changed
- 2022-01-25 17:41:34
@article{9e2a5348-8239-4cf3-b7b9-cd18e15751ce,
abstract = {{Purpose: The purpose of this study was to explore the importance of local physical tissue support for homeostasis in the isolated retina. Methods: Full-thickness retinal sheets were isolated from adult porcine eyes. Retinas were cultured for 5 or 10 days using the previously established explant protocol with photoreceptors positioned against the culture membrane (porous polycarbonate) or the Müller cell endfeet and inner limiting membrane (ILM) apposed against the membrane. The explants were analyzed morphologically using hematoxylin and eosin staining, immunohistochemistry, TUNEL labeling, and transmission electron microscopy (TEM). Results: Standard cultures displayed a progressive loss of retinal lamination and extensive cell death, with activated, hypertrophic Müller cells. In contrast, explants cultured with the ILM facing the membrane displayed a maintenance of the retinal laminar architecture, and a statistically significant attenuation of photoreceptor and ganglion cell death. TEM revealed intact synapses as well as preservation of normal cellular membrane structures. Immunohistochemistry showed no signs of Müller cell activation (GFAP), with maintained expression of important metabolic markers (GS, bFGF). Conclusion: Providing physical support to the inner but not the outer retina appears to prevent the tissue collapse resulting from perturbation of the normal biomechanical milieu in the isolated retinal sheet. Using this novel paradigm, gliotic reactions are attenuated, and metabolic processes vital for tissue health are preserved which significantly increases neuronal cell survival. This finding opens up new avenues of adult retinal tissue culture research, and increases our understanding of pathological reactions in biomechanically related conditions in vivo.}},
author = {{Taylor, Linnéa and Arnér, Karin and Holmgren Taylor, Ingrid and Ghosh, Fredrik}},
issn = {{1552-5783}},
language = {{eng}},
number = {{4}},
pages = {{2200--2213}},
publisher = {{Association for Research in Vision and Ophthalmology Inc.}},
series = {{Investigative Ophthalmology & Visual Science}},
title = {{Feet on the ground: Physical support of the inner retina is a strong determinant for cell survival and structural preservation in vitro.}},
url = {{https://lup.lub.lu.se/search/files/1360753/4647252.pdf}},
doi = {{10.1167/iovs.13-13535}},
volume = {{55}},
year = {{2014}},
}