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167 results for “rpe”
Untargeted metabolomics analysis of RPE cells during six month in culture
<p>Primary RPE cell cultures were established with human fetal RPE cells acquired from ScienCell (Cat. No, 6540) seeded at passage 3 (P3) in 12-well Transwell<sup>®</sup> inserts (Corning Inc., Cat. No. CLSS3460) coated with 2% v/v Geltrex<sup>®</sup> matrix (Thermo Fisher Scientific; Cat. No. A1413202). Cells were cultured for a total time of 6 months (25 weeks), following the same protocol as previous works (16,59). Samples were acquired for protein immunolocalization, transcriptomic and metabolomics analysis from independent cell cultures along the total 6 months culture time (specifically at 4, 12, 17 and 25 weeks in culture), collecting 3 biological replicates per type of analysis and time point.<em> </em>Cell metabolites were quenched and extracted using MeOH:H<sub>2</sub>O (80:20, -20ºC) containing a spiked solution of isotopically enriched low molecular mass internal standard<strong>. </strong>Quality control (QC) samples were prepared by creating a pool of equal volumes from each biological replicate and were analyzed after a blank solution every fifth sample to monitor the performance, stability, and reproducibility of the LC-MS run. A liquid chromatography (LC) system 1290 Infinity II (Agilent Technologies) was coupled to an Agilent 6560B Ion Mobility quadrupole-time-of-flight mass spectrometer (IM-QTOF-MS) equipped with an Agilent G1607A dual jetstream ESI source and the MassHunter WorkSation 11.0. A reference solution containing purine and hexakis(1H,1H,3H-tetrafluoropropoxy)phosphazene for mass correction was added using the second ESI source. Chromatographic, ion source and MS conditions were optimized using QCs and selected parameters are shown in Supporting Information. MS analysis was conducted with an untargeted approach, operating the instrument in both positive and negative ionization modes. All samples were analysed in a randomized order. Raw data was processed using the software Profinder B10.00 (Agilent) and refined data were exported as CEF files<strong>. </strong>Data were converted to mzml files using MS convert.</p>
Somatic copy number and structural variation in RPE-1 cells with induced chromosomal instability
<p><span><span><span><span><span><span><span><span><span><span><span>The chromosome breakage-fusion-bridge (BFB) cycle is a mutational process that produces gene amplification and genome instability. Signatures of BFB cycles can be observed in cancer genomes alongside chromothripsis, another catastrophic mutational phenomenon. Here, we explain this association by elucidating a mutational cascade, downstream of <a>th</a></span></span></span></span></span></span></span></span></span></span></span><span><span><span><span><span><span><span><span><span><span><span>e single cell division error of chromosome bridge formation, that rapidly generates extreme genomic complexity. We show that actomyosin forces are required for initial bridge breakage and mutagenesis, following which chromothripsis accumulates with aberrant interphase replication of bridge DNA. This is then followed by an unexpected burst of DNA replication in the next mitosis, generating extensive DNA damage. During this second cell division, broken bridge chromosomes frequently mis-segregate and form micronuclei, promoting additional chromothripsis. We <a>fu</a></span></span></span></span></span></span></span></span></span></span></span><span><span><span><span><span><span><span><span><span><span><span>rther show that this mutational cascade generates the continuing evolution and sub-clonal heterogeneity characteristic of many human cancers.</span></span></span></span></span></span></span></span></span></span></span></p>
KerNet, 3D fluorescence and segmentation datasets of keratin networks from MDCK, HaCaT, and RPE cells
<p><span>Mechanobiology requires precise quantitative spatial information on processes taking place in specific 3D microenvironments. Connecting microscopical, molecular, biochemical and cell mechanical data with defined topologies has turned out to be extremely difficult. Establishing such structural and functional 3D maps is particular challenging for the cytoskeleton, which consists of long and interwoven filamentous polymers coordinating subcellular processes and interactions of cells with their environment. Here we present raw data and processed segmented data of fluorescence-tagged keratin intermediate filaments from MDCK, HaCaT and RPE epithelial cells. The 30 datasets contain unprocessed airyscan superresolution 3D image data and numerical network data of the 3D arrangement of the keratin intermediate filament cytoskeleton. Information is provided on network organization at the subcellular level, including mesh arrangement, density and isotropic configuration as well as details on filament morphology such as bundling, curvature and orientation. The datasets contain data in exchange format for 3d software packages. Comparison of the resulting network parameters helps to identify similarities and differences of keratin network organization in epithelial cell types. The described approach and the presented data are pivotal for generating mechanobiological models that can be experimentally tested. </span></p>
RPE-1 GFP-H2B iCas9 sgFBXW7-A cell undergoing mitosis with a lagging chromosome that forms a micronucleus
<p>Zeiss AxioObserver video showing an RPE-1 GFP-H2B iCas9 sgFBXW7-A cell undergoing mitosis with a lagging chromosome that forms a micronucleus. Cells were treated with 1 μg/mL doxycycline for 96 hours to induce Cas9 expression and were imaged every 2 minutes for 24 hours at 40x magnification. At 15:44 chromosomes begin to condense (prophase); at 16:04 chromosomes begin to align on the metaphase plate (prometaphase); at 16:10 the chromosomes are aligned (metaphase); at 16:12 chromosomes are pulled to opposite poles of the cell (anaphase) and the lagging chromosome can be seen above the separating chromosomes; at 16:18 the chromosomes begin to unravel (telophase) and the lagging chromosome can be seen on the left-hand side of the chromosomes of the left daughter cell; from 16:28 onwards the lagging chromosome forms a micronucleus next to the nucleus of the daughter cell.</p>
RPE-1 GFP-H2B iCas9 cells undergoing mitosis
<p>Zeiss AxioObserver video showing RPE-1 GFP-H2B iCas9 cells undergoing mitosis. Cells were treated with 1 μg/mL doxycycline for 96 hours to induce Cas9 expression and were imaged every 2 minutes for 24 hours at 40x magnification.</p>
RPE-1 iCas9 sgNT cells undergoing nocodazole-induced mitotic arrest
<p>Phasefocus LiveCyte video showing RPE-1 iCas9 sgNT cells undergoing nocodazole-induced mitotic arrest. Cells were treated with 1 μg/mL doxycycline for 96 hours to induce Cas9 expression and were imaged every 9 minutes for 48 hours at 20x magnification. From 1:11 cells start entering mitosis as indicated by their rounding up into a ball; from 9:35 mitotic cells begin to slip from mitosis as indicated by rounded mitotic cells flattening. A mitotic cell undergoes mitotic cell death at 42:35 as indicated by the presence of propidium iodide (red) in the top right corner.</p>
RPE-1 GFP-H2B iCas9 cells undergoing nocodazole-induced mitotic arrest
<p>Phasefocus LiveCyte video showing RPE-1 GFP-H2B iCas9 cells undergoing nocodazole-induced mitotic arrest. Cells were treated with 1 μg/mL doxycycline for 96 hours to induce Cas9 expression and were imaged every 9 minutes for 48 hours at 20x magnification. Mitotic cells can be seen from 0:00 as indicated by their rounding up into a ball; from 2:41 mitotic cells begin to slip from mitosis as indicated by rounded mitotic cells flattening. A dead cell can be seen from 0:00 as indicated by the presence of propidium iodide (red) and two other mitotic cells undergo mitotic cell death at 43:42 and 44:20 in the bottom left corner.</p>
RPE-1 GFP-H2B iCas9 cells undergoing paclitaxel-induced mitotic arrest
<p>Phasefocus LiveCyte video showing RPE-1 GFP-H2B iCas9 cells undergoing paclitaxel-induced mitotic arrest. Cells were treated with 1 μg/mL doxycycline for 96 hours to induce Cas9 expression and were imaged every 9 minutes for 48 hours at 20x magnification. From 3:17 cells start entering mitosis as indicated by their rounding up into a ball; from 12:44 mitotic cells begin to either slip from mitosis as indicated by rounded mitotic cells flattening or undergoing mitotic cell death as indicated by the presence of propidium iodide (red).</p>
Cirrus HD-OCT Measurement of Area of Increased Light Penetration Under the Retinal Pigment Epithelium (RPE)
ClinicalTrials.gov study NCT01272076. IPD Sharing: Not stated. Countries: 1. Publications: 6.
KerNet, 3D fluorescence and segmentation datasets of keratin networks from MDCK, HaCaT, and RPE cells
Open the record for dataset details and reuse information.
Somatic copy number and structural variation in RPE-1 cells with induced chromosomal instability
Open the record for dataset details and reuse information.
Data from: Oxidative stress in the retina and retinal pigment epithelium (RPE): role of aging, and DJ-1
<p>High levels of oxidative radicals generated by daily light exposure and high metabolic rate suggest that the antioxidant machinery of the retina and retinal pigment epithelium (RPE) is crucial for their survival. DJ-1 is a redox-sensitive protein that has been shown to have neuroprotective function in the brain in Parkinson's disease and other neurodegenerative diseases. Here, we analyzed the role of DJ-1 in the retina during oxidative stress and aging. We induced low-level oxidative stress in young (3-month-old) and old (15-month-old) C57BL/6J (WT) and DJ-1 knockout (KO) mice and evaluated effects in the RPE and retina. Absence of DJ-1 resulted in increased retinal dysfunction in response to low levels of oxidative stress. Our findings suggest that loss of DJ-1 affects the RPE antioxidant machinery, rendering it unable to combat and neutralize low-level oxidative stress, irrespective of age. Moreover, they draw a parallel to the retinal degeneration observed in AMD, where the occurrence of genetic variants may leave the retina and RPE unable to fight sustained, low-levels of oxidative stress.</p>
Long Term Follow Up of Sub-retinal Transplantation of hESC Derived RPE Cells in Stargardt Macular Dystrophy Patients
ClinicalTrials.gov study NCT02445612. IPD Sharing: NO. Countries: 1. Publications: 2.
Long Term Follow Up of Sub-retinal Transplantation of hESC Derived RPE Cells in Patients With AMD
ClinicalTrials.gov study NCT02463344. IPD Sharing: NO. Countries: 1. Publications: 2.
Safety & Efficacy of Eyecyte-RPE™ in Patients With Geographic Atrophy Secondary to Dry Age-related Macular Degeneration.
ClinicalTrials.gov study NCT06394232. IPD Sharing: NO. Countries: 1. Publications: 1.
Safety and Tolerability of Sub-retinal Transplantation of Human Embryonic Stem Cell Derived Retinal Pigmented Epithelial (hESC-RPE) Cells in Patients With Stargardt's Macular Dystrophy (SMD)
ClinicalTrials.gov study NCT01469832. IPD Sharing: NO. Countries: 1. Publications: 3.
Sub-retinal Transplantation of hESC Derived RPE(MA09-hRPE)Cells in Patients With Stargardt's Macular Dystrophy
ClinicalTrials.gov study NCT01345006. IPD Sharing: NO. Countries: 1. Publications: 3.
Transplantation of Autologous RPE Versus Translocation of Autologous RPE and Choroid in AMD
ClinicalTrials.gov study NCT00401713. IPD Sharing: Not stated. Countries: 1. Publications: 2.
Safety and Tolerability of Sub-retinal Transplantation of hESC Derived RPE (MA09-hRPE) Cells in Patients With Advanced Dry Age Related Macular Degeneration
ClinicalTrials.gov study NCT01344993. IPD Sharing: NO. Countries: 1. Publications: 2.
Measurement and Analysis of Macular Retinal Pigment Epithelium (RPE) Elevations With Cirrus HD-OCT vs. Color Fundus Photography
ClinicalTrials.gov study NCT01272063. IPD Sharing: Not stated. Countries: 1. Publications: 9.
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Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
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DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.