Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe

Aline Stedman (1, 2) , Antonin Levy (3, 2, 4) , Philippe Sansonetti (5, 6, 2) , Giulia Nigro (2, 7)

1 IBPS - Institut de Biologie Paris Seine
2 Pathogénie Microbienne Moléculaire
3 Département de radiothérapie [Gustave Roussy]
4 RaMo-IT - Radiothérapie Moléculaire et Innovation Thérapeutique
5 IPS-CAS - Institut Pasteur de Shanghai, Académie des Sciences de Chine - Chinese Academy of Sciences
6 CdF (institution) - Collège de France
7 Microenvironnement et Immunité - Microenvironment and Immunity

The present protocol describes a method to detect reactive oxygen species (ROS) in the intestinal murine organoids using qualitative imaging and quantitative cytometry assays. This work can be potentially extended to other fluorescent probes to test the effect of selected compounds on ROS. ABSTRACT: Reactive oxygen species (ROS) play essential roles in intestinal homeostasis. ROS are natural byproducts of cell metabolism. They are produced in response to infection or injury at the mucosal level as they are involved in antimicrobial responses and wound healing. They are also critical secondary messengers, regulating several pathways, including cell growth and differentiation. On the other hand, excessive ROS levels lead to oxidative stress, which can be deleterious for cells

Mots clés: Murine intestinal organoids; ROS detection; Flow cytometry analysis; Live imaging detection; intestinal stem cells; ROS-sensitive dye; oxidative stress; fluorogenic probes; Murine intestinal organoids

Domaines: Biologie cellulaire; Sciences du Vivant [q-bio]

Aline Stedman, Antonin Levy, Philippe Sansonetti, Giulia Nigro. Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe. Journal of visualized experiments : JoVE, 2021, 175, ⟨10.3791/62880⟩. ⟨pasteur-03584354⟩