Derivation and Characterization of Murine and Amphibian Müller Glia Cell Lines

• DOI: 10.1167/tvst.11.4.4
• Publication Date: 2022-04-04T16:30:56Z
• Authors: Ryan A. Gallo; Farhan Qureshi; Thomas A. Strong; Steven H. Lang; Kevin A. Pino; Galina Dvoriantch...; Daniel Pelaez
• Keywords: Gliosis; Muller glia; Purinergic Signalling

Purpose: Müller glia (MG) in the retina of Xenopus laevis (African clawed frog) reprogram to a transiently amplifying retinal progenitor state after an injury. These progenitors then give rise new neurons. In contrast, mammalian MG have restricted neurogenic capacity and undergo reactive gliosis This study sought establish cell lines from regeneration-competent frog regeneration-deficient mouse. Methods: were isolated postnatal day 5 GLAST-CreERT; Rbfl/fl mice adult (3–5 years post-metamorphic) X laevis. Serial adherent subculture resulted spontaneously immortalized cells establishment two lines: murine 17 (RG17) 69 (XG69). They characterized for gene protein expression by qPCR, immunostaining, Western blot. Purinergic signaling was assessed with calcium imaging. Pharmacological perturbations 2'-3'-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) KN-62 performed on RG17 cells. Results: XG69 express several markers retain purinergic signaling. intracellular responses BzATP suggest that ionotropic receptor P2X7 is present functional Stimulation triphosphate–induced dose-dependent manner. Conclusions: We report characterization XG69, novel species significantly disparate regenerative capabilities. Translational Relevance: line models will aid comparative studies between endowed varied facilitate development cell-based strategies treating degenerative diseases.