Quantification of Reactive Oxygen Species Using 2′,7′- Dichlorofluorescein Diacetate Probe and Flow-Cytometry in Müller Glial Cells

VAGLIENTI, MARÍA V.; SUBIRADA, PAULA V.; BARCELONA, PABLO F.; BONACCI, GUSTAVO; SANCHEZ, MARÍA C.

Journal of Visualized Experiments

Journal of Visualized Experiments

Año: 2022 vol. 2022

1940-087X

he redox balance has an important role in maintaining cellular homeostasis. The increased generation of reactive oxygen species (ROS) promotes the modification of proteins, lipids, and DNA, which finally may lead to alteration in cellular function and cell death. Therefore, it is beneficial for cells to increase their antioxidant defense in response to detrimental insults, either by activating an antioxidant pathway like Keap1/Nrf2 or by improving redox scavengers (vitamins A, C, and E, β-carotene,and polyphenols, among others). Inflammation and oxidative stress are involved in the pathogenesis and progression of retinopathies, such as diabetic retinopathy (DR) and retinopathy of prematurity (ROP). Since Müller glial cells (MGCs) play a key role in the homeostasis of neural retinal tissue, they are considered an ideal model to study these cellular protective mechanisms. In this sense, quantifying ROS levels with a reproducible and simple method is essential to assess the contribu