Effects of Glial Cell Line-derived Neurotrophic Factor on the Cultured Adult Full-thickness Porcine Retina

Taylor, Linnéa; Arnér, Karin; Engelsberg, Karl; Ghosh, Fredrik

doi:10.3109/02713683.2013.763989

Cited by 27 publications

(25 citation statements)

References 34 publications

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“…In retinal explant cultures under standard conditions there are several well-characterized reactions easily observable as early as 3 or 4 DIV [32][33][34]. These reactions include gliosis and neuroretinal degeneration and can be visualized by GFAP upregulation, disruption of the cell layers and the labeling of apoptotic cells.…”

Section: The In Vitro Modelmentioning

confidence: 99%

A new model for in vitro testing of vitreous substitute candidates

Barth

Crafoord

O’Shea

et al. 2014

Graefes Arch Clin Exp Ophthalmol

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Purpose To describe a new model for in vitro assessment of novel vitreous substitute candidates. Methods The biological impact of three vitreous substitute candidates was explored in a retinal explant culture model; a polyalkylimide hydrogel (Bio-Alcamid®), a two component hydrogel of 20 wt.% poly (ethylene glycol) in phosphate buffered saline (PEG) and a cross-linked sodium hyaluronic acid hydrogel (Healaflow®). The gels where applied to explanted adult rat retinas and then kept in culture for 2, 5 and 10 days. Gel-exposed explants were compared with explants incubated under standard tissue culture conditions. Cryosections of the specimens were stained with hematoxylin and eosin, immunohistochemical markers (GFAP, Vimentin, Neurofilament 160, PKC, Rhodopsin) and TUNEL. Results Explants kept under standard conditions as well as PEG-exposed explants displayed disruption of retinal layers with moderate pyknosis of all neurons. They also displayed moderate labeling of apoptotic cells. Bio-Alcamid®-exposed explants displayed severe thinning and disruption of retinal layers with massive cell death. Healaflow®-treated explants displayed normal retinal lamination with significantly better preservation of retinal neurons compared with control specimens, and almost no signs of apoptosis. Retinas exposed to Healaflow® and retinas kept under standard conditions showed variable labeling of GFAP with generally low expression and some areas of upregulation. PEG-exposed retinas showed increased GFAP labeling and Bio-Alcamid®-exposed retinas showed sparse labeling of GFAP. Conclusions Research into novel vitreous substitutes has important implications for both medical and surgical vitreoretinal disease. The in vitro model presented here provides a method of biocompatibility testing prior to more costly and cumbersome in vivo experiments. The explant culture system imposes reactions within the retina including disruption of layers, cell death and gliosis, and the progression of these reactions can be used for comparison of vitreous substitute candidates. Bio-Alcamid® had strong adverse effects on the retina which is consistent with results of prior in vivo trials. PEG gel elicits reactions similar to the control retinas whereas Healaflow® shows protection from culture-induced trauma indicating favorable biocompatibility.

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Section: The In Vitro Modelmentioning

confidence: 99%

A new model for in vitro testing of vitreous substitute candidates

Barth

Crafoord

O’Shea

et al. 2014

Graefes Arch Clin Exp Ophthalmol

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“…Isolated adult retinal sheets cultured under standard conditions often display gliosis and neuronal degeneration over time. The discrepancy of cell survival in vitro, depending on the stage of maturity of the retinal tissue, is well established in the literature but not yet fully understood [Fernandez-Bueno et al, 2008;Kaempf et al, 2008;Kobuch et al, 2008;Taylor et al, 2013a]. Nevertheless, retina explant cultures have been widely used to study neurodegeneration, neuroprotection, and various applications [Hatakeyama and Kageyama, 2002;Manabe et al, 2002;Zhang et al, 2002;Donovan and Dyer, 2006;Leaver et al, 2006;Koizumi et al, 2007;Xin et al, 2007;Bermel et al, 2009;Feigenspan et al, 2010].…”

Section: Ex Vivo Retina Explant Culturesmentioning

confidence: 99%

“…The TUNEL assay has been used extensively for the detection and quantification of apoptosis in histological sections of retina explants [Kaempf et al, 2008;Kobuch et al, 2008;Taylor et al, 2013aTaylor et al, , 2013bTaylor et al, , 2014Barth et al, 2014;Mollick et al, 2016;Müller et al, 2017;Schnichels et al, 2017]. However, the interpretation and specificity of this assay have been controversial since TUNEL staining can identify DNA fragmentation, a characteristic of both apoptotic as well as necrotic cells [Charriaut-Marlangue and Ben-Ari, 1995;Grasl-Kraupp et al, 1995].…”

Section: Apoptosis In Retina Explantsmentioning

confidence: 99%

“…A more recent study also treated retina explants from C57BL/6 mice with Z-VAD-FMK for up to 7 days in culture, and then examined RGC density and caspase-3 activity relative to a vehicle control . Taylor et al [2013a] explored the potential neuroprotective effects of glial cell line-derived neurotrophic factor (GDNF) on cultured porcine neuroretina, focusing on the survival of each retinal cell type. The retina explants were maintained for up to 10 days ex vivo in culture medium supplemented with 10 ng/mL GDNF.…”

Section: Drug Discovery and Development Using Ex Vivo Neuroretinamentioning

confidence: 99%

“…Most retina explants are prepared as frozen sections, where the samples are fixed in 4% paraformaldehyde and cryoprotected via immersion in graded sucrose solutions Taylor et al, 2013aTaylor et al, , 2013bTaylor et al, , 2014Müller et al, 2017]. Sections > 10 µm are cut with a cryotome and stained.…”

Section: Retinal Microarchitecture and Morphometric Analysismentioning

confidence: 99%

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Current Advancements in the Development and Characterization of Full-Thickness Adult Neuroretina Organotypic Culture Systems

Rettinger

Wang²

2018

Cells Tissues Organs

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Retinal degenerative diseases such as macular degeneration, glaucoma, and diabetic retinopathy constitute the leading cause of blindness in the industrialized world. There is a continuous demand in investigative ophthalmic research for the development of new treatment modalities for retinal therapy. Unfortunately, efforts to identify novel neuroprotective and neuroregenerative agents have often been hindered by an experimental model gap that exists between high-throughput methods via dissociated cells and preclinical animal models. Even though dissociated cell culture is rapid and high-throughput, it is limited in its ability to reproduce the in vivo conditions. In contrast, preclinical animal models may offer greater fidelity, albeit they lack efficiency and experimental control. Retina explant cultures provide an ideal bridge to close this gap and have been used to study an array of biological processes such as retinal development and neurodegeneration. However, it is often difficult to interpret findings from these studies due to the wide variety of experimental species and culture methods used. This review provides a comprehensive overview of current ex vivo neuroretina culture methods and assessments, with a focus on their suitability, advantages, and disadvantages, along with novel insights and perspectives on the organotypic culture model as a high-throughput platform for screening promising molecules for retinal regeneration.

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Seeing through the interface: poly(ε -Caprolactone) surface modification of poly(glycerol-co-sebacic acid) membranes in adult porcine retinal explants

Taylor

Arnér

Kolewe

et al. 2016

J Tissue Eng Regen Med

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The purpose of this study was to investigate the adhesion properties and tissue reactions in an in vitro model of nanofabricated membranes emulating the vitreous cortex. Electrospinning was performed for either 5, 10 or 15 min to create various thicknesses of poly(ε-caprolactone) (PCL) fibre mats on a poly(glycerol-co-sebacic acid) (PGS) surface. These were fused with adult porcine retinal explants, with the fibre side facing the inner retina, and cultured for 5 days. Adherence was assessed by macroscopic inspection, and morphological and immunohistochemical analysis was performed using haematoxylin and eosin (H&E) and markers for photoreceptors and Müller glia (recoverin, NeuN, vimentin and GFAP). TUNEL labelling was performed to assess apoptosis. Five minute specimens displayed poor adherence with an overall structure, apoptosis and photoreceptor and ganglion cell morphology comparable to that of the culture controls, whereas 10 min specimens showed improved neuronal survival; 15 min composite explants adhered only at focal points, were thin and showed extensive degenerative damage. The physical composition of nanofibre meshes is important for adhesion to the inner retina and has a significant impact on neuronal and glial survival in vitro. The results bearing on research involving retinal transplantation are discussed.

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Effects of Glial Cell Line-derived Neurotrophic Factor on the Cultured Adult Full-thickness Porcine Retina

Cited by 27 publications

References 34 publications

A new model for in vitro testing of vitreous substitute candidates

A new model for in vitro testing of vitreous substitute candidates

Current Advancements in the Development and Characterization of Full-Thickness Adult Neuroretina Organotypic Culture Systems

Seeing through the interface: poly(ε -Caprolactone) surface modification of poly(glycerol-co-sebacic acid) membranes in adult porcine retinal explants

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