| Autor: |
Gloria Astolfi, Carmen Ciavarella, Sabrina Valente, Chiara Coslovi, Danilo Iannetta, Luigi Fontana, Gianandrea Pasquinelli, Piera Versura |
| Přispěvatelé: |
Astolfi, Gloria, Ciavarella, Carmen, Valente, Sabrina, Coslovi, Chiara, Iannetta, Danilo, Fontana, Luigi, Pasquinelli, Gianandrea, Versura, Piera |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
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| Popis: |
The aim of this study was to optimize a coculture in vitro model established between the human Muller glial cells and human umbilical vein endothelial cells, mimicking the inner blood-retinal barrier, and to explore its resistance to damage induced by oxidative stress. A spontaneously immortalized human Muller cell line MIO-M1 and human umbilical vein endothelial cells (HUVEC) were plated together at a density ratio 1:1 and maintained up to the 8th passage (p8). The MIO-M1/HUVECs p1 through p8 were treated with increasing concentrations (range 200-800 mu M) of H2O2 to evaluate oxidative stress induced damage and comparing data with single cell cultures. The following features were assayed p1 through p8: doubling time maintenance, cell viability using MTS assay, ultrastructure of cell-cell contacts, immunofluorescence for Vimentin and GFAP, molecular biology (q-PCR) for GFAP and CD31 mRNA. MIO-M1/HUVECs cocultures maintained distinct cell cytotype up to p8 as shown by flow cytometry analysis, without evidence of cross activation, displaying cell-cell tight junctions mimicking those found in human retina, only acquiring a slight resistance to oxidative stress induction over the passages. This MIO-M1/HUVECs coculture represents a simple, reproducible and affordable model for in vitro studies on oxidative stress-induced retinal damages. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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