Other Publications (1)
Articles by Levon Djenderedjian in JoVE
Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry Sumana R. Chintalapudi1, Need N. Patel1, Zachary K. Goldsmith1, Levon Djenderedjian1, Xiang Di Wang1, Tony N. Marion2, Monica M. Jablonski1,3,4, Vanessa M. Morales-Tirado1,2 1Department of Ophthalmology, Hamilton Eye Institute, University of Tennessee Health Science Center, 2Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, 3Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, 4Department of Pharmaceutical Sciences, University of Tennessee Health Science Center Millions of people suffer from retinal degenerative diseases that result in irreversible blindness. A common element of many of these diseases is the loss of retinal ganglion cells (RGCs). This detailed protocol describes the isolation of primary murine RGCs by positive and negative selection with flow cytometry.
Other articles by Levon Djenderedjian on PubMed
Isolation and Molecular Profiling of Primary Mouse Retinal Ganglion Cells: Comparison of Phenotypes from Healthy and Glaucomatous Retinas Frontiers in Aging Neuroscience. 2016 | Pubmed ID: 27242509 Loss of functional retinal ganglion cells (RGC) is an element of retinal degeneration that is poorly understood. This is in part due to the lack of a reliable and validated protocol for the isolation of primary RGCs. Here we optimize a feasible, reproducible, standardized flow cytometry-based protocol for the isolation and enrichment of homogeneous RGC with the Thy1.2(hi)CD48(neg)CD15(neg)CD57(neg) surface phenotype. A three-step validation process was performed by: (1) genomic profiling of 25-genes associated with retinal cells; (2) intracellular labeling of homogeneous sorted cells for the intracellular RGC-markers SNCG, brain-specific homeobox/POU domain protein 3A (BRN3A), TUJ1, and RNA-binding protein with multiple splicing (RBPMS); and (3) by applying the methodology on RGC from a mouse model with elevated intraocular pressure (IOP) and optic nerve damage. Use of primary RGC cultures will allow for future careful assessment of important cell specific pathways in RGC to provide mechanistic insights into the declining of visual acuity in aged populations and those suffering from retinal neurodegenerative diseases.