Cellular and Cytoskeletal Dynamics Within Organ Cultures of Porcine Neuroretina

Winkler, Jörg; Hagelstein, Sebastian; Rohde, Manfred; Laqua, H.

doi:10.1006/exer.2002.1188

Cited by 39 publications

(45 citation statements)

References 29 publications

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“…[10][11][12][13][14] To simulate loss of retinal tension in vitro, retinal explants can be isolated from its surrounding tissues and from the influence of the IOP. 13 Traditionally, retinal tissue is maintained in standard tissue culture dishes with artificial cerebrospinal fluid media and minimal tissue support provided by a culture membrane. Adult retina under these conditions shows poor survival, with widespread neuronal cell death after 3 to 4 days.…”

Section: Discussionmentioning

confidence: 99%

“…13 Like in detachment in vivo, activated Müller cells upregulate intermediary filaments, inducing stiffening of the tissue as well as release of cytokines and intracellular stores of glutamate. 24,28,29 These effects already lead to profound neuronal cell death after 3 DIV, and rapid formation of a gliotic scar, phenomena that are also seen in vivo after prolonged retinal detachment.…”

Section: Figure 6 Immunohistochemical Labeling Of Müller Cells Usingmentioning

confidence: 99%

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Stretch to See: Lateral Tension Strongly Determines Cell Survival in Long-Term Cultures of Adult Porcine Retina

Taylor

Moran

Arnér

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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PURPOSE.To explore the effect of lateral tension as a survival factor for retinal explants in vitro. The central nervous system (CNS) resides in a highly mechanical milieu. However, the importance of biomechanical homeostasis for normal CNS function has not been extensively explored. Diseases in which normal mechanical forces are disrupted, such as retinal detachment of the eye, are highly debilitating and the mechanisms underlying disease progression are not fully understood.METHODS. Using a porcine animal model, we developed a novel technique of culturing adult retinal explants under stretch for up to 10 days in vitro (DIV). These were compared with standard (no stretch) and free-floating cultured explants. Cell survival was analyzed using immunohistochemistry, and retinal architecture using hematoxylin and eosin staining.RESULTS. Compared with unstretched specimens, which at 10 DIV degenerated into a gliotic cell mass, stretched retinas displayed a profound preservation of the laminar retinal architecture as well as significantly increased neuronal cell survival, with no signs of impending gliosis. CONCLUSIONS.The results confirm that biomechanical tension is a vital factor in the maintenance of retinal tissue integrity, and suggest that mechanical cues are important components of pathologic responses within the CNS. (Invest Ophthalmol Vis

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Section: Discussionmentioning

confidence: 99%

Section: Figure 6 Immunohistochemical Labeling Of Müller Cells Usingmentioning

confidence: 99%

Stretch to See: Lateral Tension Strongly Determines Cell Survival in Long-Term Cultures of Adult Porcine Retina

Taylor

Moran

Arnér

et al. 2013

Invest. Ophthalmol. Vis. Sci.

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“…[13][14][15][16][17] A coronal section of the globe was made and the cornea, lens, and vitreous tissues were removed as one piece. The retina was carefully dislodged, and the remaining traces of retinal tissue were removed from the optic nerve with a sterile scalpel blade.…”

Section: Cell Isolation and Culturementioning

confidence: 99%

Conditioned medium from mixed retinal pigmented epithelium and Muller cell cultures reduces in vitro permeability of retinal vascular endothelial cells

Tretiach¹

2004

British Journal of Ophthalmology

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Aim: To investigate the in vitro effect of laser photocoagulation on blood-retinal barrier permeability. Methods: Retinal capillary endothelial cells were exposed to supernatants from long term co-cultured cells that were argon laser treated. Endothelial cell permeability was analysed by (1) measurement of transendothelial electrical resistance and (2) Results: Laser photocoagulation of various retinal cells and control ECV304 cells in the lower chamber did not appreciably improve permeability of the endothelial cell monolayer compared with that of unlasered cells. However, medium that was conditioned by mixed retinal pigmented epithelium and Müller cells significantly reduced both inulin (43.2% (SD 6.5%) equilibration in mixed cultures v 59.8% (SD 7.0%) control cells, p,0.05) and albumin (15.1% (SD 3.8%) v 31.1% (SD 6.7%), p,0.05) permeability of the endothelial cell monolayers. A fourfold increase in transendothelial electrical resistance was also seen. Conclusions: These results are consistent with the hypothesis that interaction of Müller cells with retinal pigmented epithelium induced by laser treatment results in secretion of soluble factor(s), which reduces permeability of retinal vascular endothelium. Identification of these factor(s) may have implications for the clinical treatment of macular oedema secondary to diabetic retinopathy and other diseases.

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“…Loss of of these stabilizing forces is associated with a rapid gliotic response and neuronal cell death, both in explant cultures and diseases such as retinal detachment [3,4]. Our group has previously demonstrated that in vitro, these reactions can be attenuated by simulating conditions in the normal eye, either through laterally stretching retinal explants during culture, or providing the explants with physical support at the inner border [5].…”

Section: Introductionmentioning

confidence: 99%

“…Mitochondrial DNA (mtDNA) is also particularly prone to oxidative damage [15] making the mitochondria-rich retina vulnerable to oxidative damage. 4 Normally, ROS and other oxidants are counteracted by anti-oxidants including the high-molecular weight enzymes heme oxygenase-1 (HO-1), superoxide dismutase (SOD), catalase, and glutathione peroxidases and the low-molecular weight non-enzymatic compounds glutathione, vitamin C and E. These compounds are well-characterized, but novel antioxidants are continuously being discovered. One such molecule is the reductase, hemeand radical-binding protein A1M (α1-microglobulin), which was shown to have protective properties against oxidative stress in cell cultures, skin and placenta [16][17][18], and antioxidative therapeutic effects in vivo [19][20][21] (reviewed in [22;23]).…”

Section: Introductionmentioning

confidence: 99%

The Role of Mitochondria, Oxidative Stress, and the Radical-binding Protein A1M in Cultured Porcine Retina

Åkerström

Cederlund

Bergwik

et al. 2017

Current Eye Research

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Purpose: The purpose of this study was to explore the relationship between oxidative stress, antioxidant defense, mitochondrial structure and biomechanical tissue support in the isolated porcine retina. Methods:Full-thickness retinal sheets were isolated from adult porcine eyes. Retinas were cultured for 2 or 48 hours using 1) a previously established low-support explant protocol with photoreceptors positioned against the culture membrane (porous polycarbonate) or 2) a high-support procedure developed by our group, apposing the Müller cell endfeet and inner limiting membrane (ILM) against the membrane. The grafts were analyzed by quantitative PCR, immunohistochemistry, and transmission electron microscopy (TEM), and culture medium was assayed for the cell damage and oxidative stress markers lactate dehydrogenase and protein carbonyls. Results:In explants cultured with physical support to the inner border, cone photoreceptors were preserved and lactate dehydrogenase levels were reduced, although an initial (2h), transient, increased oxidative stress was observed. Elevated expression of the antioxidants A1M (α1-microglobulin) and heme oxygenase-1 were seen in the mitochondria-rich inner segments after 48h compared to low-support counterparts.Housekeeping gene expression suggested a higher degree of structural integrity of mitochondria in high-support explants, and TEM of inner segments confirmed preservation of a normal mitochondrial morphology. Conclusion:Providing retinal explants with inner retinal support leads to mobilization of antioxidant proteins, preservation of mitochondrial function and increased cell viability.Consequently, the failure of low-support retinal cultures to mobilize an adequate response to 2 the oxidative environment may play an key role in their rapid demise. These findings shed new light on pathological reactions in biomechanically related conditions in vivo.

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Cellular and Cytoskeletal Dynamics Within Organ Cultures of Porcine Neuroretina

Cited by 39 publications

References 29 publications

Stretch to See: Lateral Tension Strongly Determines Cell Survival in Long-Term Cultures of Adult Porcine Retina

Stretch to See: Lateral Tension Strongly Determines Cell Survival in Long-Term Cultures of Adult Porcine Retina

Conditioned medium from mixed retinal pigmented epithelium and Muller cell cultures reduces in vitro permeability of retinal vascular endothelial cells

The Role of Mitochondria, Oxidative Stress, and the Radical-binding Protein A1M in Cultured Porcine Retina

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