Journal: Molecular Vision

Article Title: Physical disruption of cell–cell contact induces VEGF expression in RPE cells

doi:

Figure Lengend Snippet: ICC results for scratched cultures of ARPE-19 and hRPE cells. Results show confocal images for short-term (1 day after confluency) and long-term (4 weeks after confluency) cultures of ARPE-19 ( A - C , E - G ) and hRPE ( I - K , M - O ) cells. VEGF expression was elevated along the edges of scratched areas in all sample types. Graphs D , H , L , and P show net signal intensity for VEGF along the length of the relevant confocal image (line graphs) and the average VEGF signal intensity at 0-100 μm and 100-200 μm from the scratched edge (bar graphs). The net signal intensity was higher proximal to the scratch zone and lower in confluent areas for all samples. The average signal intensity was higher in the first 100 μm from the scratch edge for all samples. Blue = 4',6-diamidino-2-phenylindole (DAPI); red = VEGF. Yellow lines indicate the scratch edge. Scale bars = 100 μm.

Article Snippet: The Certification of Analysis of the hRPE from Lonza is also included as passages 1–4 of the hRPE cells were used to generate the results found here.

Techniques: Expressing

Journal: Molecular Vision

Article Title: Physical disruption of cell–cell contact induces VEGF expression in RPE cells

doi:

Figure Lengend Snippet: ICC results for confluent hRPE cultures. Cells were immunostained for Claudin-1 ( A , B ), N-cadherin ( C , D ), β-catenin ( E , F ) and ZO-1 ( G , H ). I - L : Percentage of intercellular junctions covered by the corresponding protein in cultures of hRPE cells grown for 1 day or 4 weeks after reaching confluency. All junctional proteins, except for ZO-1, had limited localization after 1 day post confluence. In all cases, junctional localization increased markedly after 4 weeks. Data represent the mean ± standard deviation for three replicates from three representative confocal images per each time point for each junctional protein (n = 3). * p<0.05, ** p<0.01. M . Z-Stack scan for a long-term (4 weeks after confluency) culture of hRPE cells grown on porous cell culture inserts, confirming apical localization of ZO-1 and basolateral localization of VEGF. Scale bar = 25 μm.

Article Snippet: The Certification of Analysis of the hRPE from Lonza is also included as passages 1–4 of the hRPE cells were used to generate the results found here.

Techniques: Standard Deviation, Cell Culture

Journal: Molecular Vision

Article Title: Physical disruption of cell–cell contact induces VEGF expression in RPE cells

doi:

Figure Lengend Snippet: Micropatterning of ARPE-19 and hRPE cells. A – C : Scanning electron microscope (SEM) images of through holes of different sizes ( A : 100, B : 200, and C : 300 μm in diameter) in the polydimethylsiloxane (PDMS) stencils. The lateral spacing between the holes was 1 mm for all pattern sizes. D – F : ARPE-19 cells attached to glass coverslips through PDMS stencil holes after 24 h. G – I : hRPE cells on glass coverslips, after the PDMS stencils were removed. Scale bars, A – C = 500 μm; D – I = 200 μm.

Article Snippet: The Certification of Analysis of the hRPE from Lonza is also included as passages 1–4 of the hRPE cells were used to generate the results found here.

Techniques: Microscopy

Journal: Molecular Vision

Article Title: Physical disruption of cell–cell contact induces VEGF expression in RPE cells

doi:

Figure Lengend Snippet: Micropatterning results. A : Formation of varying concentrations of ARPE-19 cell with a free edge by micropatterning. Black = average number of cells per cell patch of corresponding size; red = concentration of cells losing cell–cell contact at least on one side. Increase in the concentration of cells losing cell–cell contact correlates negatively with cell count and patch size. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). Similar results were seen for the human RPE (hRPE) cells (data not shown). B : Increase in the cell patch diameter at day 2. A greater increase in patch diameter was observed for the ARPE-19 cells compared with the hRPE cells. Data represent the mean ± standard deviation for three replicates (seven measurements per replicate for each pattern size and each time point; n = 21). C , D : VEGF expression analysis with enzyme-linked immunosorbent assay (ELISA) for the micropatterned ARPE-19 ( C ) and hRPE ( D ) cells. VEGF expression is presented as the total VEGF divided by the cell count. The smaller the pattern size, the higher the VEGF expression per cell. VEGF expression decreased during day 2 presumably because of the increased pattern sizes due to cell growth ( B ). Data represent the mean ± standard deviation for three replicates (three measurements per replicate for each pattern size and each time point; n = 9); * p<0.05, ** p<0.01, *** p<0.001. P values are relative to the control (confluent) samples.

Article Snippet: The Certification of Analysis of the hRPE from Lonza is also included as passages 1–4 of the hRPE cells were used to generate the results found here.

Techniques: Concentration Assay, Cell Counting, Standard Deviation, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Molecular and cellular therapies

Article Title: Neural stem cells: ready for therapeutic applications?

doi: 10.1186/2052-8426-2-31

Figure Lengend Snippet: Clinical trials involving stem cells for retinal dystrophies

Article Snippet: NCT01625559 , Safety and Tolerability of MA09-hRPE Cells in Patients With Stargardt’s Macular Dystrophy (SMD) , Recruiting , Stargardt’s Macular Dystrophy , Biological: MA09-hRPE implantation , CHA Bio & Diostech , Phase 1 , 3 , September 2012 , October 2014.

Techniques: Clinical Proteomics, Transplantation Assay, Derivative Assay

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: DEL-1 ameliorates ER stress in hRPE cells. A Western blot analysis depicting phosphorylated eIF2α and CHOP in hRPE cells treated with tunicamycin (1 μg/mL) and/or DEL-1 (0–1 μg/mL) for 24 h. B Western blot analysis depicting phosphorylated eIF2α and CHOP in hRPE cells treated with DEL-1 (0–2 μg/mL) for 24 h. Means ± SDs were calculated from three independent experiments in in vitro models. * P < 0.05 compared with the control. ! P < 0.05 compared with tunicamycin treatment

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Western Blot, In Vitro, Control

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: DEL-1 mitigates ER stress-induced VEGF expression and apoptosis in hRPE cells. A Cell viability assay in hRPE cells treated with tunicamycin (0–2 μg/mL) or DEL-1 (0–2 μg/mL) for 24 h. B Western blot analysis of VEGF in hRPE cells treated with tunicamycin (1 μg/mL) and/or DEL-1 (0–1 μg/mL) for 24 h. C Cell viability and caspase 3 activity assay in hRPE cells treated with tunicamycin (2 μg/mL) and/or DEL-1 (0–5 μg/mL) for 24 h. Means ± SDs were calculated from three independent experiments in in vitro models. * P < 0.05 compared with the control. ! P < 0.05 compared with tunicamycin (1 or 2 μg/mL) treatment

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Expressing, Viability Assay, Western Blot, Caspase-3 Activity Assay, In Vitro, Control

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: Involvement of AMPK in the effects of DEL-1 on ER stress, VEGF expression, and apoptosis in hRPE cells. A Western blot analysis of phosphorylated AMPK, CAMKK2 and LKB1 in hRPE cells treated with DEL-1 (0–1 μg/mL) for 24 h. B Western blot analysis of phosphorylated eIF2α, CHOP, and VEGF in AMPK siRNA-transfected hRPE cells treated with tunicamycin (1 μg/mL) and/or DEL-1 (1 μg/mL) for 24 h. C Cell viability and caspase 3 activity assay in AMPK siRNA-transfected hRPE cells treated with tunicamycin (2 μg/mL) and/or DEL-1 (1 μg/mL) for 24 h. Means ± SDs were calculated from three independent experiments in in vitro models. * P < 0.05 compared with the control. ! P < 0.05 compared with tunicamycin (1 or 2 μg/mL) treatment. # P < 0.05 compared with tunicamycin (1 or 2 μg/mL) and DEL-1 treatment

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Expressing, Western Blot, Transfection, Caspase-3 Activity Assay, In Vitro, Control

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: Autophagy-mediated signaling contributes to the protective effects of DEL-1 against ER stress-induced VEGF expression and injury in hRPE cells. A MDC staining and Western blot analysis of LC3 I/II and p62 in hRPE cells treated with DEL-1 (0–1 μg/mL) for 24 h. B Western blot analysis of phosphorylated eIF2α, CHOP, and VEGF in hRPE cells treated with tunicamycin (1 μg/mL), DEL-1 (1 μg/mL), and/or 3-MA (2 mM) for 24 h. C Cell viability and caspase 3 activity assay in hRPE cells treated with tunicamycin (1 μg/mL), DEL-1 (1 μg/mL), and/or 3-MA (2 mM) for 24 h. Means ± SDs were calculated from three independent experiments in in vitro models. * P < 0.05 compared with the control. ! P < 0.05 compared with tunicamycin (1 or 2 μg/mL) treatment. # P < 0.05 compared with tunicamycin (1 or 2 μg/mL) and DEL-1 treatment

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Expressing, Staining, Western Blot, Caspase-3 Activity Assay, In Vitro, Control

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: DEL-1 attenuates ER stress and apoptosis in primary hRPE cells. A Western blot analysis depicting phosphorylated eIF2α and CHOP in primary hRPE cells treated with tunicamycin (1 μg/mL) and/or DEL-1 (0–1 μg/mL) for 24 h. B Cell viability and caspase 3 activity assay in primary hRPE cells treated with tunicamycin (2 μg/mL) and/or DEL-1 (0–1 μg/mL) for 24 h. Means ± SDs were calculated from three independent experiments in in vitro models. * P < 0.05 compared with the control. ! P < 0.05 compared with tunicamycin (1 or 2 μg/mL) treatment

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Western Blot, Caspase-3 Activity Assay, In Vitro, Control

Journal: Journal of Translational Medicine

Article Title: DEL-1: a promising treatment for AMD-associated ER stress in retinal pigment epithelial cells

doi: 10.1186/s12967-024-04858-9

Figure Lengend Snippet: Illustration depicting the effects of the myokine DEL-1 on VEGF expression and apoptosis in hRPE cells

Article Snippet: Cultivation of primary hRPE cells obtained from Sciencell Research Laboratories (San Diego, CA, USA) was performed using optimized Epithelial Cell Medium (Sciencell Research Laboratories).

Techniques: Expressing