NDLI: Differential expression of neuronal genes in Müller glia in two- and three-dimensional cultures.

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Differential expression of neuronal genes in Müller glia in two- and three-dimensional cultures.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Phillips, M. Joseph Otteson, Deborah C.
Description	Country affiliation: United States Author Affiliation: Phillips MJ ( Department of Basic Sciences, College of Optometry, University of Houston, Houston, Texas 77204-2020, USA.)
Abstract	PURPOSE: Müller glia in the mammalian retina have some stem cell-like characteristics, although their capacity for neurogenesis remains limited both in vivo and in vitro. In vitro studies to date have used traditional two-dimensional (2D) cell culture to assess neuronal differentiation of Müller glia. The purpose of this study was to compare the effects of 2D and three-dimensional (3D) environments on Müller glial gene expression after growth factor stimulation. METHODS: Conditionally immortalized mouse Müller glia cells (ImM10) were cultured under nonimmortalizing conditions with EGF/FGF2 to generate spheres that were differentiated in vitro on uncoated culture dishes (2D) or encapsulated in self-assembling, RADA-16 peptide hydrogels (3D) under identical media and growth factor supplementation conditions. Gene expression was analyzed using quantitative RT-PCR and immunocytochemistry. Cellular morphology was analyzed with light and confocal microscopy; sphere ultrastructure was analyzed with transmission electron microscopy. RESULTS: ImM10 Müller cells express numerous genes associated with neural stem cells and retinal progenitors in both normal growth conditions and sphere-forming conditions. When encapsulated in the 3D hydrogel, cells can migrate and send processes into the hydrogel. Many genes associated with neurogenesis, as well as retinal neuron-specific genes, are differentially expressed in 2D and 3D differentiation conditions. CONCLUSIONS: ImM10 Müller glia upregulate genes characteristic of retinal neurons after growth factor stimulation in vitro, and gene expression patterns are altered in 3D hydrogel cultures.
ISSN	01460404
e-ISSN	15525783
DOI	10.1167/iovs.10-6400
Journal	Investigative Opthalmology & Visual Science
Issue Number	3
Volume Number	52
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2011-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Cell Differentiation Physiology Gene Expression Regulation Nerve Tissue Proteins Genetics Neurogenesis Neuroglia Cytology Retinal Neurons Animals Cell Culture Techniques Epidermal Growth Factor Pharmacology Fibroblast Growth Factor 2 Immunohistochemistry Mice Mice, Inbred C57bl Microscopy, Confocal Microscopy, Electron, Transmission Neural Stem Cells Metabolism Drug Effects Rna, Messenger Reverse Transcriptase Polymerase Chain Reaction Spheroids, Cellular Ultrastructure Tissue Scaffolds Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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