Characterization of Retinal Pigment Epithelial Melanin and Degraded Synthetic Melanin Using Mass Spectrometry and In Vitro Biochemical Diagnostics

2018 ◽  
Vol 95 (1) ◽  
pp. 183-191 ◽  
Author(s):  
Sally M. Yacout ◽  
Kelsey L. McIlwain ◽  
Shama P. Mirza ◽  
Elizabeth R. Gaillard
Keyword(s):  
Mass Spectrometry ◽  
Retinal Pigment Epithelial ◽  
Retinal Pigment ◽  
Pigment Epithelial

scholarly journals A Safe GDNF and GDNF/BDNF Controlled Delivery System Improves Migration in Human Retinal Pigment Epithelial Cells and Survival in Retinal Ganglion Cells: Potential Usefulness in Degenerative Retinal Pathologies

2021 ◽  
Vol 14 (1) ◽  
pp. 50
Author(s):  
Alicia Arranz-Romera ◽  
Maria Hernandez ◽  
Patricia Checa-Casalengua ◽  
Alfredo Garcia-Layana ◽  
Irene T. Molina-Martinez ◽  
...  
Keyword(s):  
Cell Viability ◽  
Retinal Ganglion Cells ◽  
Neurotrophic Factor ◽  
Retinal Pigment Epithelial ◽  
Ganglion Cells ◽  
Retinal Pigment ◽  
Terminal Deoxynucleotidyl Transferase ◽  
Retinal Ganglion ◽  
Pigment Epithelial

We assessed the sustained delivery effect of poly (lactic-co-glycolic) acid (PLGA)/vitamin E (VitE) microspheres (MSs) loaded with glial cell-derived neurotrophic factor (GDNF) alone (GDNF-MSs) or combined with brain-derived neurotrophic factor (BDNF; GDNF/BDNF-MSs) on migration of the human adult retinal pigment epithelial cell-line-19 (ARPE-19) cells, primate choroidal endothelial (RF/6A) cells, and the survival of isolated mouse retinal ganglion cells (RGCs). The morphology of the MSs, particle size, and encapsulation efficiencies of the active substances were evaluated. In vitro release, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell viability, terminal deoxynucleotidyl transferase (TdT) deoxyuridine dUTP nick-end labelling (TUNEL) apoptosis, functional wound healing migration (ARPE-19; migration), and (RF/6A; angiogenesis) assays were conducted. The safety of MS intravitreal injection was assessed using hematoxylin and eosin, neuronal nuclei (NeuN) immunolabeling, and TUNEL assays, and RGC in vitro survival was analyzed. MSs delivered GDNF and co-delivered GDNF/BDNF in a sustained manner over 77 days. The BDNF/GDNF combination increased RPE cell migration, whereas no effect was observed on RF/6A. MSs did not alter cell viability, apoptosis was absent in vitro, and RGCs survived in vitro for seven weeks. In mice, retinal toxicity and apoptosis was absent in histologic sections. This delivery strategy could be useful as a potential co-therapy in retinal degenerations and glaucoma, in line with future personalized long-term intravitreal treatment as different amounts (doses) of microparticles can be administered according to patients’ needs.

Cyanidin-3-glucoside attenuates 4-hydroxynonenal- and visible light-induced retinal damage in vitro and in vivo

2019 ◽  
Vol 10 (5) ◽  
pp. 2871-2880 ◽  
Author(s):  
Yong Wang ◽  
Wentao Qi ◽  
Yazhen Huo ◽  
Ge Song ◽  
Hui Sun ◽  
...  
Keyword(s):  
Retinal Pigment Epithelial ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Retinal Damage ◽  
Protective Effects ◽  
Pigment Epithelial ◽  
Induced Apoptosis ◽  
Pigment Epithelial Cells

Cyanidin-3-glucoside has efficient protective effects on 4-hydroxynonenal-induced apoptosis, senescence, and angiogenesis in retinal pigment epithelial cells.

Effect of hrWnt7a on Human Retinal Pigment Epithelial Cells In Vitro

2015 ◽  
Vol 159 (4) ◽  
pp. 534-540 ◽  
Author(s):  
A. V. Kuznetsova ◽  
A. M. Kurinov ◽  
E. V. Chentsova ◽  
P. V. Makarov ◽  
M. A. Aleksandrova
Keyword(s):  
Epithelial Cells ◽  
Retinal Pigment Epithelial ◽  
Retinal Pigment ◽  
Retinal Pigment Epithelial Cells ◽  
Pigment Epithelial ◽  
Pigment Epithelial Cells

scholarly journals Synchronization of human retinal pigment epithelial-1 cells in mitosis

2020 ◽  
Vol 133 (18) ◽  
pp. jcs247940
Author(s):  
Stacey J. Scott ◽  
Kethan S. Suvarna ◽  
Pier Paolo D'Avino
Keyword(s):  
Cell Lines ◽  
Large Scale ◽  
Retinal Pigment Epithelial ◽  
Retinal Pigment ◽  
Interaction Network ◽  
Protein Protein Interaction ◽  
Pigment Epithelial ◽  
Adenocarcinoma Cells ◽  
Sequential Treatments

ABSTRACTHuman retinal pigment epithelial-1 (RPE-1) cells are increasingly being used as a model to study mitosis because they represent a non-transformed alternative to cancer cell lines, such as HeLa cervical adenocarcinoma cells. However, the lack of an efficient method to synchronize RPE-1 cells in mitosis precludes their application for large-scale biochemical and proteomics assays. Here, we report a protocol to synchronize RPE-1 cells based on sequential treatments with the Cdk4 and Cdk6 inhibitor PD 0332991 (palbociclib) and the microtubule-depolymerizing drug nocodazole. With this method, the vast majority (80–90%) of RPE-1 cells arrested at prometaphase and exited mitosis synchronously after release from nocodazole. Moreover, the cells fully recovered and re-entered the cell cycle after the palbociclib–nocodazole block. Finally, we show that this protocol could be successfully employed for the characterization of the protein–protein interaction network of the kinetochore protein Ndc80 by immunoprecipitation coupled with mass spectrometry. This synchronization method significantly expands the versatility and applicability of RPE-1 cells to the study of cell division and might be applied to other cell lines that do not respond to treatments with DNA synthesis inhibitors.

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