TY - JOUR. T1 - Repigmentation of human retinal pigment epithelial cells in vitro. AU - Boulton, Mike. AU - Marshall, John. PY - 1985/8. Y1 - 1985/8. N2 - Cultured human retinal pigment epithelial (RPE) cells readily ingested both melanin and lipofuscin isolated from human RPE cells. Up to 7 days post-challenge ingested granules demonstrated no evidence of lysis or aggregation within secondary lysosomes. When cultures containing ingested melanin and lipofuscin were subcultured the cells gradually depigmented due to a redistribution of pigment granules amongst daughter cells. Quantitative analysis demonstrated that the accumulation of both types of granule was linear over a 24 hr challenge period. This study reports a technique of artificially repigmenting cultured human RPE cells and thus offers the potential for in vitro investigations of the role of these inclusions in various dynamic aspects of cellular metabolism.. AB - Cultured human retinal pigment epithelial (RPE) cells readily ingested ...
TY - JOUR. T1 - Genotoxic effects of carotenoid breakdown products in human retinal pigment epithelial cells. AU - Kalariya, Nilesh M.. AU - Ramana, Kota. AU - Srivastava, Satish. AU - Van Kuijk, Frederik J G M. PY - 2009. Y1 - 2009. N2 - Purpose: To investigate the genotoxic effects of lutein (LBP) and β -carotene breakdown products (β -apo-8-carotenal, BA8C) and the preventive role of GSH in human retinal pigment epithelial cells (ARPE-19). Methods: LBP- and BA8C-induced DNA damage in human retinal pigment epithelial cells (ARPE-19) was determined by comet assay. The DNA damage was quantified by the image analysis system using Comet Score™ software. ARPE-19 cell viability was determined by CellTiter 96 AQueous one-solution cell proliferation assay kit. Intracellular GSH levels were measured by Ellmans reagent. Results: Incubation of serum-starved ARPE-19 cells with LBP and BA8C caused significant DNA damage in a dose- and time-dependent manner. The DNA damage and cell death incurred by ...
PURPOSE: To characterize glutathione (GSH) transport by cultured human retinal pigment epithelial (HRPE) cells. METHODS: Cultured HRPE cells were pretreated with acivicin for GSH efflux and with buthionine sulfoximine for GSH uptake to prevent the breakdown and resynthesis of GSH. Efflux was measured by the linear rate of accumulation of GSH in the supernatant; uptake was measured using [35S] GSH plus varying concentrations of GSH. Molecular forms were verified by high-performance liquid chromatography. HRPE cell mRNA was probed for the presence of the two recently cloned rat sinusoidal and canalicular GSH transporters, (RsGshT and RcGshT), by Northern blot analysis. RESULTS: Glutathione efflux was temperature dependent (undetectable at 4 degrees C), and its averaged 23 +/- 3.3 pmol/10(6) cells/minute or 10% of the total GSH effluxed per hour (total cell GSH = 13.6 +/- 1.5 nmol/10(6) cells). Efflux was not influenced by dithiothreitol or sulfobromophthalein-reduced GSH adduct, agents known to ...
Purpose: : To investigate the in vitro effects of tobacco-specific n- nitrosamines (TSNA); (N-Nitrosonornicotine (NNN), 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) which are the major components of cigarette smoke on human retinal pigment epithelial cells. Methods: : Human ARPE-19 cultures were exposed to varying concentrations of NNN and NNK (1mM, 100µM, 10µM, 1 µM) in DMEM/F12 medium for 24 and 48 hours. MTT cell proliferation assays were performed to find a non toxic dose that would not kill the cells within the 24-48 hour period. For both NNN and NNK a 10µM dose of 24 hours was chosen to carry out the rest of the experiments. Total cell protein extracts were analyzed for the presence of OGG1, a DNA repair enzyme and VEGF. Live cells were stained with Hoechst and Mitotracker Green to visualize the nucleus and mitochondrial mass respectively using a confocal microscope. Cells were also labeled for anti- VEGFR2 and imaged using a confocal microscope. Comet assay was performed on ...
The goal of the present study was to determine whether treatment with cigarette smoke extract (CSE) induces cell loss, cellular senescence, and extracellular matrix (ECM) synthesis in primary human retinal pigment epithelial (RPE) cells. Primary cultured human RPE cells were exposed to 2, 4, 8, and 12% of CSE concentration for 24 hours. Cell loss was detected by cell viability assay. Lipid peroxidation was assessed by loss of cis-parinaric acid (PNA) fluorescence. Senescence-associated ß-galactosidase (SA-ß-Gal) activity was detected by histochemical staining. Expression of apolipoprotein J (Apo J), connective tissue growth factor (CTGF), fibronectin, and laminin were examined by real-time PCR, western blot, or ELISA experiments. The results showed that exposure of cells to 12% of CSE concentration induced cell death, while treatment of cells with 2, 4, and 8% CSE increased lipid peroxidation. Exposure to 8% of CSE markedly increased the number of SA-ß-Gal positive cells to up to 82%, and the mRNA
TY - JOUR. T1 - L-carnitine protects human retinal pigment epithelial cells from oxidative damage. AU - Shamsi, Farrukh A.. AU - Chaudhry, Imtiaz A.. AU - Boulton, Mike E.. AU - Al-Rajhi, Ali A.. PY - 2007/6/1. Y1 - 2007/6/1. N2 - Purpose: To determine the efficacy of L-carnitine (LC) against oxidative changes in human retinal pigment epithelium (RPE) cells. Methods: The RPE cells from human donor eyes were cultured in Hams F-10 medium. The effect of LC on H2O2-induced morphologic changes in the RPE cells was analyzed by light microscopy. Reduction in cell death after the impact of LC treatment on H2O2-treated cells was analyzed by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide] assays. In addition, the effect of H2O2 on the activity of RPE-antioxidant enzymes, glutathione (GSH) and superoxide dismutase (SOD), and LC-induced protection was also determined. Results: LC protected the RPE cells by inhibiting the peroxide-induced cytopathic effect from 50% to 10%. Nuclear ...
TY - JOUR. T1 - Proliferation of retinal pigment epithelial cells induced by (R, R)-XY-10 and (S, S)-XY-10 and their action mechanisms. AU - Cheng, Yu Wen. AU - Wang, Yu Liang. AU - Zhang, Yi Hua. AU - Peng, Si Xun. AU - Chiou, George C.Y.. PY - 2009/10/22. Y1 - 2009/10/22. N2 - • AIM: To investigate the mechanism of proliferation effect induced by (R, R)-XY-10 and (S, S)-XY-10 on retinal pigmented epithelial cells (ARPE-19). • METHODS: Human retinal pigmented epithelial cells (ARPE-19) and human umbilical vein endothelial cells (HUVECs) were used to investigate the effect of (R, R)-XY-10 and (S, S)-XY-10 on cell growth, and their mechanisms of proliferative action by using ERKinverted commas AKTinverted commas PI3Kinverted commas Protein kinase C (PKC)and Nitric oxide synthase (NOS) inhibitors. RESULTS: (R, R)-XY-10 and (S, S)-XY-10 dose-dependently increased ARPE-19 cell proliferation, but not on HUVECs. When treated with proliferative inhibitors, H7 (5μ mol/L)inverted commas hypericin ...
TY - JOUR. T1 - Age-related susceptibility to apoptosis in human retinal pigment epithelial cells is triggered by disruption of p53-Mdm2 association.. AU - Bhattacharya, Sujoy. AU - Chaum, Edward. AU - Johnson, Dianna A.. AU - Johnson, Leonard R.. PY - 2012/1/1. Y1 - 2012/1/1. N2 - Relatively little is known about the contribution of p53/Mdm2 pathway in apoptosis of retinal pigment epithelial (RPE) cells or its possible link to dysfunction of aging RPE or to related blinding disorders such as age-related macular degeneration (AMD). Age-associated changes in p53 activation were evaluated in primary RPE cultures from human donor eyes of various ages. Apoptosis was evaluated by activation of caspases and DNA fragmentation. Gene-specific small interfering RNA was used to knock down expression of p53. We observed that the basal rate of p53-dependent apoptosis increased in an age-dependent manner in human RPE. The age-dependent increase in apoptosis was linked to alterations in several aspects of the ...
In the present study, we investigated the effects of blue light filtering on the secretion profile of proangiogenic cytokines by RPE cells. To the best of our knowledge, this is the first study to demonstrate that following light exposure, angiogenin secretion by RPE cells is decreased, and this decrease is abrogated with a blue light filter. We also demonstrated that, although not significant, this trend is maintained when RPE cells are grown under hypoxic conditions and when pre-treated with lutein.. Although the etiology of AMD is not well understood, there is strong evidence indicating that retinal hypoxia plays a significant role in retinal neovascularization [21, 22]. On the other hand, lutein-containing supplements have been found to be beneficial and protective in slowing the progression of AMD [23-26]. We therefore sought to determine whether the effects of blue light filtering are maintained when RPE cells are exposed to hypoxic conditions or when pre-treated with lutein. We focused ...
Choroidal neovascularization (CNV) is the most severe form of age-related macular degeneration (AMD), which causes rapid visual loss. Transplantation of cultured retinal pigment epithelium (RPE) cell sheet by tissue engineering is a possible approach
We showed previously that confluent ARPE-19 cells grown on different matrices had different transcriptional profiles from native RPE, and that cells grown on plastic had the closest transcriptome to native RPE.2 To improve the transcriptional proximity of ARPE-19 cells grown on plastic to native RPE, we varied the culture conditions and evaluated global expression trends, a more informative benchmark than individual gene expression. With this end point, CSW and DSW cells were most similar to native RPE. While each culture condition preserved the expression of the most abundant genes, significant transcriptional differences exist between native and cultured cells including the number of genes on the array that were expressed and not expressed, the number and function of differentially expressed genes, and the number of expressed low abundance genes. Our results suggest that the global expression profiles can be improved by varying the culture conditions, but significant widespread differences ...
Methods In three primary RPE cell cultures (from three donor eyes) and in the human RPE cell line ARPE-19, FcRn and beta-2-microglobulin (β2M) mRNA levels were determined by real-time quantitative PCR. FcRn protein expression was analysed by western blot studies. Stimulation experiments were performed with recombinant human tumour necrosis factor (TNF)-α and interferon (IFN)-γ. HT-29, THP-1 and HeLa cell lines were used as FcRn positive and negative non-ocular controls, respectively. ...
Research resource: nuclear receptor atlas of human retinal pigment epithelial cells: potential relevance to age-related macular degeneration.
The retinal pigment epithelium (RPE) is constantly exposed to external injuries which lead to degeneration, dysfunction or loss of RPE cells. The balance between RPE cells death and proliferation may be responsible for several diseases of the underlying retina, including age-related macular degeneration (AMD) and proliferative vitreoretinopathy (PVR). Signaling pathways able to control cells proliferation or death usually involve the MAPK (mitogen-activated protein kinases) pathways, which modulate the activity of transcription factors by phosphorylation. UV exposure induces DNA breakdown and causes cellular damage through the production of reactive oxygen species (ROS) leading to programmed cell death. In this study, human retinal pigment epithelial cells ARPE19 were exposed to 100 J/m(2) stop of UV-C and MAPK pathways were studied. We first showed the expression of the three major MAPK pathways. Then we showed that activator protein-1 (AP-1) was activated through phosphorylation of cJun and ...
Dept of Biochemistry, Cell Biology was started in a small way in the year 1985 . Active research was started in 1991 by Prof. S. Ramakrishnan and Dr. K. N. Sulochana into Ocular Biochemistry. The extensive areas of research includes, cataract, Eales disease, Diabetic Retinopathy, pathways of homocysteine and proteoglycan, novel protein isolation, use of amino acid as antiglycating substances to mitigate diabetic complication were the focus of the lab.. Later Dr. N. Angayarkanni since 2002 has been working in primary cultures of Bovine retinal pigment epithelium, retinal capillary Endothelial and pericytes focussing on ECM changes in the vitreo-retinal pathologies apart from establisbing proteomics. Current Focus is on drug development for the treatment of ocular angiogenesis, alterations in metabolic pathways leading to diseases, structural function studies of proteins, protein-protein interactions, clinical proteomics, lipidomics, toxicology ...
Retinal pigment epithelial cells (RPE) stably expressed human CXCR4 when transduced with N-terminal FLAG or N-terminal FLAG and C-terminal MYC
TY - JOUR. T1 - Ascorbate suppresses VEGF expression in retinal pigment epithelial cells. AU - Sant, David W.. AU - Camarena, Vladimir. AU - Mustafi, Sushmita. AU - Li, Yiwen. AU - Wilkes, Zachary. AU - van Booven, Derek. AU - Wen, Rong. AU - Wang, Gaofeng. PY - 2018/7/1. Y1 - 2018/7/1. N2 - PURPOSE. To investigate the impact of ascorbate, via DNA hydroxymethylation, on VEGF expression in retinal pigment epithelial (RPE) cells. METHODS. Dot-blot and hydroxymethylated DNA immunoprecipitation sequencing were applied to evaluate the impact of ascorbate on DNA hydroxymethylation in ARPE-19 cells. RNA sequencing (RNA-seq) was carried out to analyze the transcriptome. Quantitative RT-PCR and ELISA were conducted to examine the transcription and secretion of VEGF from cultured cells. Primary human fetal RPE cells and RPE-J cells were used to verify the effect of ascorbate on VEGF expression. ELISA was used to measure VEGF in the vitreous humor of Gulo-/-mice, which, like humans, cannot synthesize ...
ATCC hTERT immortalized RPE cell lines have an extended lifespan, undergo terminal differentiation, express RPE associated proteins, and are karyotypically, morphologically, and phenotypically similar to the primary parent cells.
ATCC hTERT immortalized RPE cell lines have an extended lifespan, undergo terminal differentiation, express RPE associated proteins, and are karyotypically, morphologically, and phenotypically similar to the primary parent cells.
Age-related macular degeneration (AMD) is a leading cause of worldwide blindness in the elderly. It is a bilateral ocular condition that impairs the central retina known as the macula. The macula accounts for the majority of daytime, color vision in humans. Thus, lesions in the macula have a major impact on human vision. Previous studies have suggested that oxidative stress to certain ocular cells may contribute to the development of AMD. Oxidative stress occurs when reactive oxygen species (ROS) interact with protein and DNA to modify their functions. In this study, Aryan et al used hydrogen peroxide, a highly reactive compound, to induce oxidative stress in human retinal pigment epithelial cells, a type of ocular cell which provides nourishment for the human retina. Oxidative stress resulted in a profound influence on advancing the senescence (functional deterioration) of these cells and inhibiting their proliferation. These results strongly suggest that oxidative stress plays a role in the ...
Hyperspectral AF images (hypercubes) were captured from 66, 40X fields in 11 RPE/BrM flat mounts from human donor eyes using techniques described in detail in the abstract submitted by K. Agarwal. Briefly, for each 40X field, the hypercube has the two spatial dimensions of the field, and at each spatial point the photon counts recorded at each wavelength, hence the third or spectral dimension. For reproducible quantification of these data, exposure times were calibrated so that photon counts per spectral channel fell within the 12-bit linear range of the detector and then were offset by the dark current. Scaled counts-per-second were determined by exposure time (Eqn. 1) and calibrated to a standard fluorescent reference (courtesy of F Delori) to correct for any variation in power of the excitation light, yielding quantified hypercubes with units of photon counts per second at each point and wavelength.. Results ...
In silico pathway, gene ontology, and system-level network comparisons of EBOV-infected and mock-infected ARPE-19 cell transcriptomic profiles all revealed that EBOV-infected human retinal pigment epithelial cells generated a robust type I IFN response. Consistent with these results, 28% of significantly upregulated transcripts were identified as type I IFN-responsive in the Interferome database of IFN-regulated genes.27 Although the type I IFN response is a critical innate immune defense against viral infection, this result was quite unexpected; EBOV causes severe and accelerated pathology, because it prevents the type I IFN response in other human host cells, including the mononuclear phagocyte populations that are its early targets.37 The major type I IFNs are IFN-α and IFN-β: IFN-β may be more important for the response to EBOV, because treatment with recombinant IFN-β, but not IFN-α, prolongs survival of infected macaques.38 Both IFN-α and IFN-β signal via the IFN-α/β receptor, ...
In silico pathway, gene ontology, and system-level network comparisons of EBOV-infected and mock-infected ARPE-19 cell transcriptomic profiles all revealed that EBOV-infected human retinal pigment epithelial cells generated a robust type I IFN response. Consistent with these results, 28% of significantly upregulated transcripts were identified as type I IFN-responsive in the Interferome database of IFN-regulated genes.27 Although the type I IFN response is a critical innate immune defense against viral infection, this result was quite unexpected; EBOV causes severe and accelerated pathology, because it prevents the type I IFN response in other human host cells, including the mononuclear phagocyte populations that are its early targets.37 The major type I IFNs are IFN-α and IFN-β: IFN-β may be more important for the response to EBOV, because treatment with recombinant IFN-β, but not IFN-α, prolongs survival of infected macaques.38 Both IFN-α and IFN-β signal via the IFN-α/β receptor, ...
PURPOSE: To locate the mildest and/or earliest changes in the retina and/or choroid in Sveinsson chorioretinal atrophy (SCRA), using more advanced techniques than previous studies. METHODS: We used fundus photography, intravenous fluorescein angiography (IVFA) enhanced ocular coherence tomography (OCT) scans, microperimetry and multifocal electroretinography (mfERG) in an attempt to locate the mildest changes in SCRA. Eight patients with SCRA were examined. To improve the resolution of OCT scans, several consecutive recorded B-scans were retrieved for each location of interest. The scans were processed off-line with an averaging algorithm developed to maximally reduce laser speckle (noise). Static microperimetry was performed using the Rodenstock scanning laser ophthalmoscope (SLO). RESULTS: Biomicroscopy and fundus photographs disclosed an apparent thinning of the retinal pigment epithelium (RPE) in the areas minimally affected, where possible changes in the transparent sensory retina were not ...
The selective damage of the retinal pigment epithelium (RPE) is a new treatment method for several retinal diseases. By applying a train of microsecond(s) laser pulses it is possible to selectively damage these cells and simultaneously spare the adjacent photoreceptor and neural tissue. Due to the ophthalmologic invisibility of the RPE cell damage we investigate an optoacoustic (OA) control system to monitor the RPE cell damage. Setup: The irradiation was performed with a frequency doubled Nd:YLF laser by applying a train of +s laser pulses. In vitro, the OA transients were received by an ultrasonic broadband transducer. During treatment an OA contact lens with embedded transducer was used. In vitro: Freshly enucleated porcine RPE samples with CalceinAM as life/death staining were used. Below RPE cell damage threshold a classic thermoelastic transient was found. Above cell damage threshold the OA transient differs form pulse to pulse. This can be explained by microbubble formation around the ...
The retinal pigment epithelium (RPE)1 is a highly specialized derivative of the embryonic neural tube that lies with its apical surface in intimate contact with the light-sensitive cells of the retina (Zinn and Marmor, 1979), performing critical transport, barrier, and phagocytic support functions for the neural retina. These functions of RPE cells require a characteristic apical distribution of certain proteins that are usually found on the basolateral membrane in other epithelia. For example, apical Na,K-ATPase provides a high Na+ environment appropriate for photoreceptor function (Bok, 1982; Okami et al., 1990; Gundersen et al., 1991; Gallemore et al., 1997; Miller and Steinberg, 1977; Rizzolo, 1997; Zhao et al., 1997). The apical localization of the neural cell adhesion molecule N-CAM-140 in RPE (Gundersen et al., 1993), which overrides a dominant basolateral signal in the cytoplasmic domain recognized by MDCK cells (Powell et al., 1991; Le Gall et al., 1997), is presumably required to ...
BACKGROUND: Adenoma of the retinal pigment epithelium (RPE) is a rare intraocular tumor that can simulate other pigmented tumors such as choroidal melanoma. We report a case of non-pigmented adenoma of the RPE initially diagnosed as choroidal hemangioma. CASE REPORT: A 42-year-old woman presented to Kurume University Hospital in November 1992 with an orange-yellow tumor nasal to the optic disc in the left fundus. The tumor was 9.0 × 9.0 mm in diameter, 6.0 mm thick, and was characterized by high intensity on T1-weighted magnetic resonance imaging (MRI), low intensity on T2-weighted MRI, and enhancement on gadolinium MRI. Fluorescein angiography revealed early hypofluorescence and late hyperfluorescence of the tumor and retinal feeder vessels. By April 1996, exudate had developed around the tumor margins. The patient was treated with external beam radiation therapy (20 Gy) in July 1996, but the tumor did not diminish in size. Subsequently, she developed extensive loss of vision due to total ...
The purpose of the study is to explore the safety, tolerability and efficacy of Spheramine (cultured human retinal pigment epithelial cells on microcarriers) in Parkinsons Disease patients with advanced disease who have insufficient symptom control by optimum oral medication. Patients are randomized to receive Spheramine injections into both hemispheres or a sham surgical procedure in a ratio of 1:1. A three month pretreatment period must be completed prior to surgery. Time to endpoint is 24 months ...
Metabolic relationships between cells in the retina and retinal pigment epithelium are fundamental to retinal function, retinal disease and age-related vision loss and they may provide strategies for metabolism-based therapies.
I microRNA (miRNA) profili di staminali umane pluripotenti indotte-(iPS), cellule dellepitelio pigmentato retinico (RPE) derivati ...
Results & Discussion. Generation of subtracted bovine RPE cDNA library. We generated a subtracted bovine RPE cDNA library to identify genes specifically or predominantly expressed in mammalian RPE. Subtraction was performed between a ss circular bovine RPE cDNA library and biotinylated mRNA from bovine heart and liver. The heart and liver were chosen as driver tissues for subtraction as they are developmentally different from the posterior of the eye where RPE resides. This subtraction was expected to allow the enrichment of genes expressed in RPE, a tissue of neural origin. The subtraction method required only 2 ng of ss circular RPE cDNA library as a starting material, allowing the use of more than 1000-fold molar excess of driver mRNA to target cDNA, which helped in obtaining efficient subtraction. The subtracted ss circular library cDNA was electroporated into MC1061 E. coli cells without converting into ds DNA. Rubenstein et al. [12] reported about 100 to 1000 fold higher transformation ...
Age-related macular degeneration is a leading cause of worldwide blindness in the elderly. It is a bilateral ocular condition that impairs the central retina known as the macula. The macula accounts for the majority of daytime, color vision in humans. Thus, lesions in the macula have a major impact on human vision.
This study is a Phase I/II, open-label, non randomized, sequential, multi-center clinical trial. There will be 5 cohorts, the 4 low vision cohorts will contain 3 patients, the better vision cohort will contain 4 patients. The enrolled cohorts will be as follows:. Three AMD patients- 50,000 MA09-hRPE cells transplanted Three AMD patients- 100,000 MA09-hRPE cells transplanted Four Better Vision AMD patients- 100,000 MA09-hRPE cells transplanted Three AMD patients- 150,000 MA09-hRPE cells transplanted Three AMD patients- 200,000 MA09-hRPE cells transplanted. Patients will be enrolled sequentially, and within each cohort of 3 patients, each patients clinical course over the first 6 weeks following cell transplantation will be reviewed by an independent (DSMB) before enrollment is opened for the next 2 patients. A full safety assessment of all 3 patients in each cohort will be made by the DSMB when the 3rd patient in each cohort completes 4 weeks of follow-up, and before the first patient in the ...
Retinal pigment epithelium (RPE) cell-based gene expression studies performed under hypoxia and/or hyperglycemia show huge potential for modeling cell responses in diabetic retinopathy, retinopathy of prematurity and other retinal diseases. However, normalization of gene expression on RPE cells under those conditions has commonly been done using either GAPDH or β-actin as reference genes without any validation of their expression stability. Therefore, we aimed to establish a suitable set of reference genes for studies on RPE cells cultured under both normal culturing glucose and atmospheric oxygen tension (normoxia, 21%), under a low oxygen tension (hypoxia, 1%), under a high glucose growth medium (25 mmol/l) and under the combination of the two changed conditions above for distinct time points taking together from 24h to 7 days ...
The stem cell biotech Advanced Cell Technology (ACT) reported new, positive data in a paper in Lancet from their clinical trials using retinal pigmented epithelial cells (RPEs) made from human embryonic stem cells (hESC) for treatment of different forms of macular degeneration (MD). The paper was entitled "Human embryonic stem cell-derived retinal pigment epithelium in patients with age-related macular degeneration and Stargardts macular dystrophy: follow-up of two open-label phase 1/2 studies" with first author Steven D. Schwartz and senior author Robert Lanza, CSO of ACT. These two …Read More. ...
The stem cell biotech Advanced Cell Technology (ACT) reported new, positive data in a paper in Lancet from their clinical trials using retinal pigmented epithelial cells (RPEs) made from human embryonic stem cells (hESC) for treatment of different forms of macular degeneration (MD). […]. ...
The purpose of this study was to identify and characterize previously unknown inhibitor(s) of RPE65, which controls the rate-limiting step of the visual cycle and retinal susceptibility to light-induced degeneration. Through an unbiased screening of a bovine RPE cDNA library, we isolated FATP4, ELOVL1, PSMD13, and RDH5 as candidate negative regulators of RPE65 (Fig. 1G).. RDH5 is a stereospecific RDH that catalyzes oxidation of 11cROL to 11cRAL (Simon et al., 1995). As predicted, the reduced levels of 11cROL in the RDH5-transfected cells (Fig. 1G) was the result of oxidation of 11cROL to 11cRAL by RDH5 (Fig. 2A). We therefore conclude that RDH5 is not an inhibitor of RPE65. Although RDH5 is not an inhibitor of RPE65, our results help to explain why the isomerase activity in bovine RPE microsomes is very low. RDH5 is a membrane-bound protein. A majority of bovine RDH5 is associated with smooth ER in RPE (Simon et al., 1999), indicating that RDH5 should be present in RPE microsomes. This ...
This study will be investigating human retinal pigment epithelial cell therapy [MA09-hRPE;Advanced Cell Technology] in ptients with geographic atrophy secondary
Janice M. Burke, PhD has used cells cultured from the human retinal pigment epithelium (RPE) for many years to study basic features of the structure and function of these cells.
Principal Investigator:TAZAWA Yutaka,田沢 豊, Project Period (FY):1989 - 1991, Research Category:Grant-in-Aid for General Scientific Research (C), Research Field:Ophthalmology
Generation of bovine RPE/retina-subtracted cDNA library. Detailed description of the library will be published elsewhere (J. T. Chang, N. Della, C. Chew, S. Zhang, P. A. Campochiaro, and D. J. Zack, unpublished data). In brief, a library was constructed in Uni-ZAP XR (Strategene, La Jolla, CA) using cDNA that was generated from bovine RPE RNA; the library was in vivo excised and made single-stranded, hybridized in several rounds with an excess of biotinylated heart and liver RNA; the resulting RNA-DNA hybrids and unhybridized RNA were removed by phenol extraction after the addition of streptavidin; and the remaining unhybridized plasmid DNA was electroporated into MC1061 cells.. Fluorescent in situ hybridization. Fluorescentin situ hybridization (FISH) mapping was performed by standard methods (Lichter et al., 1990). Identical results were obtained with two independent but overlapping P1 clones. The clones were identified from high-density filters and were processed according to the suppliers ...
Heath Allison K., Morphological and Proliferative Changes that Occur in Rat Retinal Progenitor Cells Following Incubation With Retinonoic Acid and RPE-Secreted Proteins. Masters of Science (Cell Biology and Genetics), August 2006, 67 pp., 12 figures, bibliography. The principal objective of this research is to characterize virally-transformed rat retinal progenitor cells following stimulation by retinal pigment epithelial (RPE) cell secreted proteins and retinoic acid. Progenitor cells were isolated from explants of postnatal rat RPE cell in vitro. Isolated progenitor cells were cloned, analyzed by microscopy and proliferation bioassays, to determine if cell proliferation occurred. The isolated progenitor cells were analyzed for differentiation by Western blot analyses and immunocytochemistry. The rat progenitor cells cultured in RPE secreted proteins proliferated, but did not differentiate as shown by the presence of nestin and vimentin in these cells. Retinoic acid caused other progenitor cells to
We study how newly synthesised proteins and proteins taken up from the cell surface are sorted and delivered to the appropriate destination within the cell. The correct traffic of these proteins plays a critical role in regulating cell proliferation, survival and differentiation in all cells. We also study specific traffic events in cells of the eye, focusing on retinal pigment epithelial cells (RPE). Melanin pigment within the RPE is critical for eye development and protects the adult eye from light and oxidative stress. We study how pigment granules are made, how they move and how defects in these processes contribute to inherited retinal degenerations. RPE also take up and degrade spent photoreceptor segments and a gradual build-up of undigested products within the RPE occurs with age and is increased in AMD. We study the regulation of degradation of outer segments in order to understand how these processes breakdown with age and disease.. ...
This broad patent gives Advanced Cell Technology the opportunity to have a monopoly around embryonic stem cells derived from RPE cells. There are over 200 diseases and conditions which potentially be...
Sigma-Aldrich offers abstracts and full-text articles by [Kapil Bharti, Melanie Gasper, Jingxing Ou, Martha Brucato, Katharina Clore-Gronenborn, James Pickel, Heinz Arnheiter].
When DArcy Wentworth Thompsons On Growth and Form was published 100 years ago, it raised the question of how biological forms arise during development and across evolution. In light of the advances in molecular and cellular biology since then, a succinct modern view of the question states: how do genes encode geometry? Our new special issue is packed with articles that use mathematical and physical approaches to gain insights into cell and tissue patterning, morphogenesis and dynamics, and that provide a physical framework to capture these processes operating across scales.. Read the Editorial by guest editors Thomas Lecuit and L. Mahadevan, as they provide a perspective on the influence of DArcy Thompsons work and an overview of the articles in this issue.. ...
Table 1. Pattern of immunolabeling in retinas of mammalian species with the anti-chick CNTFRα antibody. Intensity of labeling was graded as intense (+++), moderate (++), weak (+), or absent (0); RPE staining was not determined (ND) in some species. In mouse RPE, nonspecific labeling could also be observed on negative control sections. Labeling was intense in PD3 and PD6 rats. Data for "Dog" are taken from from a previous study [18]. For dog and human outer nuclear layer (ONL), labeling was limited to cone soma, axon, and pedicle. For cat ONL, labeling was limited to cone soma and some cone axons. For sheep and pig ONL, labeling was limited to cone soma. Retinal pigment epithelium (RPE) pigmentation and autofluorescence precluded assessing the presence of specific labeling in pig, monkey, and human. In pig inner segments (IS), labeling was limited to the inner portion of the IS.. ...
Summary GlobalDatas clinical trial report, Retinal Pigment Epithelial (RPE) Detachment Global Clinical Trials Review, H2, 2015 provides an overview of
Introduction Numerous exogenous molecules may cause toxic chorioretinitic effects. Some agents cause disruption of the retinal pigment epithelium (RPE), while others produce vascular damage within the retina. Certain agents may also produce edema of the retina, particularly in the macular region, while other agents produce crystalline deposits in the retina from derivatives of their metabolites…
I ran across an interesting paper in PLOS One published back in March of 2012 by Parameswaran G. Sreekumar, Christine Spee, Stephen J. Ryan, Susan P. C. Cole, Ram Kannan and David R. Hinton. This manuscript looks at a mechanism of retinal pigment epithelium (RPE) cell death with notable findings identifying therapeutic targets for disorders that involve the RPE cells. The […]. ...