A Critical Analysis of the Available<i>In Vitro</i>and<i>Ex Vivo</i>Methods to Study Retinal Angiogenesis

Moleiro, Ana Filipa; Conceição, Glória; Leite‐Moreira, Adelino F.; Rocha‐Sousa, Amândio

doi:10.1155/2017/3034953

Cited by 37 publications

(30 citation statements)

References 188 publications

(279 reference statements)

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“…The main advantages of this technique are the precise evaluation of vascular sprouting from mature vessels in the adult retina and the ability to evaluate potential pro-angiogenic factors. A disadvantage of this model is the lack of blood flow, circulating endothelial progenitors, and hormonal factors that a have key role in angiogenesis [26].…”

Section: Ex Vivo Assaysmentioning

confidence: 99%

Evaluation of Angiogenesis Assays

Stryker¹,

Rajabi

Davis

et al. 2019

Biomedicines

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Angiogenesis assays allow for the evaluation of pro- or anti-angiogenic activity of endogenous or exogenous factors (stimulus or inhibitors) through investigation of their pro-or anti- proliferative, migratory, and tube formation effects on endothelial cells. To model the process of angiogenesis and the effects of biomolecules on that process, both in vitro and in vivo methods are currently used. In general, in vitro methods monitor specific stages in the angiogenesis process and are used for early evaluations, while in vivo methods more accurately simulate the living microenvironment to provide more pertinent information. We review here the current state of angiogenesis assays as well as their mechanisms, advantages, and limitations.

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Section: Ex Vivo Assaysmentioning

confidence: 99%

Evaluation of Angiogenesis Assays

Stryker¹,

Rajabi

Davis

et al. 2019

Biomedicines

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“…The limitation of this study is that an animal study was not performed. Although in vitro studies are faster and easier to perform and quantify, in vitro studies can be criticized for being very different from the natural environment [ 45 ]. Further study to investigate the role of si -SOX2 using an in vivo model would be necessary.…”

Section: Discussionmentioning

confidence: 99%

Effect of SOX2 Repression on Corneal Endothelial Cells

Hwang

Shin

2020

IJMS

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Purpose: Human corneal endothelial cells (hCECs) pump out water from the stroma and maintain the clarity of the cornea. The sex-determining region Y-box 2 (SOX2) participates in differentiation during the development of the anterior segment of the eye and is found in the periphery of wounded corneas. This study was performed to investigate the effect of SOX2 repression on hCECs. Methods: Cultured hCECs were transfected by siRNA for SOX2. The wound healing rate and cell viability were measured. The cell proliferation-associated protein level was evaluated by Western blotting and RT-PCR. The energy production and mitochondrial function were measured, and cell shape and WNT signaling were assessed. Results: Upon transfecting the cultured cells with siRNA for SOX2, the SOX2 level was reduced by 80%. The wound healing rate and viability were also reduced. Additionally, CDK1, cyclin D1, SIRT1, and ATP5B levels were reduced, and CDKN2A and pAMPK levels were increased. Mitochondrial oxidative stress and mitochondrial viability decreased, and the cell shape became elongated. Furthermore, SMAD1, SNAI1, WNT3A, and β-catenin levels were increased. Conclusion: SOX2 repression disrupts the normal metabolism of hCECs through modulating WNT signaling and mitochondrial functions.

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“…There are some obvious limitations to ex vivo retina explant culture systems, examples of which include the physical separation of the RPE from the neuroretina, the absence of a choroidal and retinal blood supply, and the axotomy of RGCs due to the dissection procedure [Moleiro et al, 2017]. Consequently, the ex vivo culture systems described here are not meant to be long-term culture solutions.…”

Section: Challenges and Limitationsmentioning

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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A Critical Analysis of the AvailableIn VitroandEx VivoMethods to Study Retinal Angiogenesis

Cited by 37 publications

References 188 publications

Evaluation of Angiogenesis Assays

Evaluation of Angiogenesis Assays

Effect of SOX2 Repression on Corneal Endothelial Cells

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

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