Induction of Differentiation by Pyruvate and DMEM in the Human Retinal Pigment Epithelium Cell Line ARPE-19

Ahmado, Ahmad; Carr, Amanda-Jayne F.; Vugler, Anthony; Semo, Ma’ayan; Gias, Carlos; Lawrence, Jean M.; Chen, Li Li; Chen, Fred K.; Turowski, Patric; Cruz, Lyndon da; Coffey, Peter

doi:10.1167/iovs.10-6374

Cited by 88 publications

(100 citation statements)

References 75 publications

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“…Furthermore, microvilli-like structures were also observed on the apical surfaces of ARPE-19 cells. According to previous reports, ARPE-19 cells rarely exhibit this morphologic characteristic under routine culture conditions [42]. Our results suggest that long-term co-cultures established on RWSF/PCL/Gt membranes maintained this development potential and also induced RPE-like cellular extensions.…”

Section: Discussionsupporting

confidence: 85%

“…PEDF, a secreted factor characteristic of RPE cells, has the key neurotrophic function and, importantly, it also inhibits retinal and choroidal neovascularization [63]. Additionally, PEDF expression is also used as an important criterion to evaluate the functionality of polarized RPE cells [42]. Based on our studies, ARPE-19 cells only exhibited polar secretion of PEDF when seeded on RWSF/PCL/Gt and RWSF/PCL membranes after 4 weeks of culture.…”

Section: Discussionmentioning

confidence: 75%

“…Pigmentation is a native RPE phenotype in vivo, and it is usually utilized as an important differentiation marker of RPE cells [42]. However, pigmentation has never occurred in ARPE-19 cells cultured on artificial substrates in other studies.…”

Section: Discussionmentioning

confidence: 99%

“…For long-term studies, cells were grown for two weeks in RPE medium. After reaching confluence, the medium was replaced by DLP medium (DMEM with 1 g/L glucose, 1% FBS and 1 mM pyruvate) [42], then maintained for more than 12 weeks. DLP medium was changed twice a week.…”

Section: Generation Of Human Rpe Cells and Electrospun Membrane (Hrpementioning

confidence: 99%

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A novel Bruch's membrane-mimetic electrospun substrate scaffold for human retinal pigment epithelium cells

et al. 2014

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a b s t r a c tVarious artificial membranes have been used as scaffolds for retinal pigment epithelium cells (RPE) for monolayer reconstruction, however, long-term cell viability and functionality are still largely unknown. This study aimed to construct an ultrathin porous nanofibrous film to mimic Bruch's membrane, and in particular to investigate human RPE cell responses to the resultant substrates. An ultrathin porous nanofibrous membrane was fabricated by using regenerated wild Antheraea pernyi silk fibroin (RWSF), polycaprolactone (PCL) and gelatin (Gt) and displayed a thickness of 3e5 mm, with a high porosity and an average fiber diameter of 166 ± 85 nm. Human RPE cells seeded on the RWSF/PCL/Gt membranes showed a higher cell growth rate (p < 0.05), and a typical expression pattern of RPE signature genes, with reduced expression of inflammatory mediators. With long-term cultivation on the substrates, RPE cells exhibited characteristic polygonal morphology and development of apical microvilli. Immunocytochemisty demonstrated RPE-specific expression profiles in cells after 12-weeks of co-culture on RWSF/PCL/Gt membranes. Interestingly, the cells on the RWSF/PCL/Gt membranes functionally secreted polarized PEDF and phagocytosed labeled porcine POS. Furthermore, RWSF/PCL/Gt membranes transplanted subsclerally exhibited excellent biocompatibility without any evidence of inflammation or rejection. In conclusion, we established a novel RWSF-based substrate for growth of RPE cells with excellent cytocompatibility in vitro and biocompatibility in vivo for potential use as a prosthetic Bruch's membrane for RPE transplantation.

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Section: Discussionsupporting

confidence: 85%

Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Generation Of Human Rpe Cells and Electrospun Membrane (Hrpementioning

confidence: 99%

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A novel Bruch's membrane-mimetic electrospun substrate scaffold for human retinal pigment epithelium cells

et al. 2014

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“…The human RPE cell line ARPE-19 21 was maintained in high-glucose DMEM supplemented with 10% FBS, 1% l-glutamine, and 1% penicillin/ streptomycin at 37°C. To obtain a differentiated RPE phenotype, 22 1-1.5 · 10 5 cell/cm 2 were plated on Matrigel-coated Transwell as above, except that FBS was reduced to 1%. Cells were allowed to differentiate for 8-10 weeks.…”

Section: Rpe Cell Linesmentioning

confidence: 99%

Enhanced Functional Integration of Human Photoreceptor Precursors into Human and Rodent Retina in anEx VivoRetinal Explant Model System

Yanai

Laver

Gregory‐Evans

et al. 2015

Tissue Engineering Part A

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Retinal disease is the major cause of irreversible blindness in developed countries. Transplantation of photoreceptor precursor cells (PPCs) derived from human embryonic stem cells (hESCs) is a promising and widely applicable approach for the treatment of these blinding conditions. Previously, it has been shown that after transplantation into the degenerating retina, the percentage of PPCs that undergo functional integration is low. The factors that inhibit PPC engraftment remain largely unknown, in part, because so many adverse factors could be at play during in vivo experiments. To advance our knowledge in overcoming potential adverse effects and optimize PPC transplantation, we have developed a novel ex vivo system. Harvested neural retina was placed directly on top of cultured retinal pigment epithelial (RPE) cells from a number of different sources. To mimic PPC transplantation into the subretinal space, hESC-derived PPCs were inserted between the retinal explant and underlying RPE. Explants cocultured with hESC-derived RPE maintained normal gross morphology and viability for up to 2 weeks, whereas the explants cultured on ARPE19 and RPE-J failed by 7 days. Furthermore, the proportion of PPCs expressing ribbon synapse-specific proteins BASSOON and RIBEYE was significantly higher when cocultured with hESC-derived RPE (20% and 10%, respectively), than when cocultured with ARPE19 (only 6% and 2%, respectively). In the presence of the synaptogenic factor thrombospondin-1 (TSP-1), the proportion of BASSOON-positive and RIBEYE-positive PPCs cocultured with hESC-derived RPE increased to *30% and 15%, respectively. These data demonstrate the utility of an ex vivo model system to define factors, such as TSP-1, which could influence integration efficiency in future in vivo experiments in models of retinal degeneration.

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CYLD Maintains Retinal Homeostasis by Deubiquitinating ENKD1 and Promoting the Phagocytosis of Photoreceptor Outer Segments

Yang,

Yu,

Yang

et al. 2024

Advanced Science

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Phagocytosis of shed photoreceptor outer segments by the retinal pigment epithelium (RPE) is essential for retinal homeostasis. Dysregulation of the phagocytotic process is associated with irreversible retinal degenerative diseases. However, the molecular mechanisms underlying the phagocytic activity of RPE cells remain elusive. In an effort to uncover proteins orchestrating retinal function, the cylindromatosis (CYLD) deubiquitinase is identified as a critical regulator of photoreceptor outer segment phagocytosis. CYLD‐deficient mice exhibit abnormal retinal structure and function. Mechanistically, CYLD interacts with enkurin domain containing protein 1 (ENKD1) and deubiquitinates ENKD1 at lysine residues K141 and K242. Deubiquitinated ENKD1 interacts with Ezrin, a membrane‐cytoskeleton linker, and stimulates the microvillar localization of Ezrin, which is essential for the phagocytic activity of RPE cells. These findings thus reveal a crucial role for the CYLD‐ENKD1‐Ezrin axis in regulating retinal homeostasis and may have important implications for the prevention and treatment of retinal degenerative diseases.

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Induction of Differentiation by Pyruvate and DMEM in the Human Retinal Pigment Epithelium Cell Line ARPE-19

Cited by 88 publications

References 75 publications

A novel Bruch's membrane-mimetic electrospun substrate scaffold for human retinal pigment epithelium cells

A novel Bruch's membrane-mimetic electrospun substrate scaffold for human retinal pigment epithelium cells

Enhanced Functional Integration of Human Photoreceptor Precursors into Human and Rodent Retina in anEx VivoRetinal Explant Model System

CYLD Maintains Retinal Homeostasis by Deubiquitinating ENKD1 and Promoting the Phagocytosis of Photoreceptor Outer Segments

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