Time course modifications in organotypic culture of human neuroretina

Fernandez-Bueno, Iván; Fernández‐Sánchez, Laura; Gayoso, Manuel J.; García-Gutiérrez, María T.; Pastor, J. Carlos; Cuenca, Nicolás

doi:10.1016/j.exer.2012.08.012

Cited by 58 publications

(52 citation statements)

References 44 publications

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“…The outer segments of the cones are greatly shortened and swollen in the detached retina, and antibodies against cone opsins now label the plasma membrane of cone cells extending to the ONL . Similar changes with swollen and truncated cone outer segments have been found in organotypic cultures of human neuroretina (Fernandez-Bueno et al, 2012) and in animal models of RP (Figs. 4CeD,F and 8A) (Garcia-Ayuso et al, 2013;MartinezNavarrete et al, 2011;Pinilla et al, 2007).…”

Section: Photoreceptor Morphology Changessupporting

confidence: 51%

“…At the electron microscopy level, atypical synapses have been found in an animal model of retinal detachment and in human retinal organotypic cultures (Fernandez-Bueno et al, 2012). Furthermore, groups of 3 synaptic ribbons without their corresponding postsynaptic elements were observed in both cases.…”

Section: Photoreceptor Morphology Changesmentioning

confidence: 96%

“…Moreover, the organization of the outer segment discs, with parallel membrane alignment under normal conditions, changes to an irregular distribution of disc membranes, a common feature of RP in animal models and human organotypic cultures Fernandez-Bueno et al, 2012).…”

Section: Photoreceptor Morphology Changesmentioning

confidence: 99%

See 2 more Smart Citations

Cellular responses following retinal injuries and therapeutic approaches for neurodegenerative diseases

Cuenca

Fernández‐Sánchez

Campello

et al. 2014

Progress in Retinal and Eye Research

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353

394

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Section: Photoreceptor Morphology Changessupporting

confidence: 51%

Section: Photoreceptor Morphology Changesmentioning

confidence: 96%

See 1 more Smart Citation

Cellular responses following retinal injuries and therapeutic approaches for neurodegenerative diseases

Cuenca

Fernández‐Sánchez

Campello

et al. 2014

Progress in Retinal and Eye Research

Self Cite

353

394

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“…Briefly, the explants are fixed in glutaraldehyde, post-fixed in osmium tetroxide prior to a graded series of ethanol and infiltration of the explants with resin, followed by curing. Ultrathin sections are counterstained with uranyl acetate and lead citrate and examined using a transmission electron microscope Kobuch et al, 2008;Fernandez-Bueno et al, 2012;Taylor et al, 2014]. Ultrastructural analysis enables the visualization of the architecture of cells at higher magnifications than can be obtained under a standard optical light microscope.…”

Section: Ultrastructural Changes In Retina Explantsmentioning

confidence: 99%

“…With TEM, cellular structures such as organelles, which allow the cell to function properly within its specific environment, can be examined at the ultrastructural level. For instance, the rapid response of rod and cone outer segments to the detachment and ultrastructural features of subretinal gliosis have been investigated in detail using TEM [Winkler et al, 2002;Kobuch et al, 2008;Fernandez-Bueno et al, 2012;Taylor et al, 2014].…”

Section: Ultrastructural Changes In Retina Explantsmentioning

confidence: 99%

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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Design of synthetic promoters for controlled expression of therapeutic genes in retinal pigment epithelial cells

Johari

Mercer

Liu

et al. 2021

Biotech & Bioengineering

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Age‐related macular degeneration (AMD) associated with dysfunction of retinal pigment epithelial (RPE) cells is the most common cause of untreatable blindness. To advance gene therapy as a viable treatment for AMD there is a need for technologies that enable controlled, RPE‐specific expression of therapeutic genes. Here we describe design, construction and testing of compact synthetic promoters with a pre‐defined transcriptional activity and RPE cell specificity. Initial comparative informatic analyses of RPE and photoreceptor (PR) cell transcriptomic data identified conserved and overrepresented transcription factor regulatory elements (TFREs, 8–19 bp) specifically associated with transcriptionally active RPE genes. Both RPE‐specific TFREs and those derived from the generically active cytomegalovirus‐immediate early (CMV‐IE) promoter were then screened in vitro to identify sequence elements able to control recombinant gene transcription in model induced pluripotent stem (iPS)‐derived and primary human RPE cells. Two libraries of heterotypic synthetic promoters varying in predicted RPE specificity and transcriptional activity were designed de novo using combinations of up to 20 discrete TFREs in series (323–602 bp) and their transcriptional activity in model RPE cells was compared to that of the endogenous BEST1 promoter (661 bp, plus an engineered derivative) and the highly active generic CMV‐IE promoter (650 bp). Synthetic promoters with a highpredicted specificity, comprised predominantly of endogenous TFREs exhibited a range of activities up to 8‐fold that of the RPE‐specific BEST1 gene promoter. Moreover, albeit at a lower predicted specificity, synthetic promoter transcriptional activity in model RPE cells was enhanced beyond that of the CMV‐IE promoter when viral elements were utilized in combination with endogenous RPE‐specific TFREs, with a reduction in promoter size of 15%. Taken together, while our data reveal an inverse relationship between synthetic promoter activity and cell‐type specificity, cell context‐specific control of recombinant gene transcriptional activity may be achievable.

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Time course modifications in organotypic culture of human neuroretina

Cited by 58 publications

References 44 publications

Cellular responses following retinal injuries and therapeutic approaches for neurodegenerative diseases

Cellular responses following retinal injuries and therapeutic approaches for neurodegenerative diseases

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

Design of synthetic promoters for controlled expression of therapeutic genes in retinal pigment epithelial cells

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