Document Detail


A conditional immortalized mouse muller glial cell line expressing glial and retinal stem cell genes.
MedLine Citation:
PMID:  20505190     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Müller glia have multiple functions in the retina, including synthesis of neurotrophic factors, uptake and metabolism of neurotransmitters, spatial buffering of ions, maintenance of the blood-retinal barrier, and response to injury. A population of Müller glia has some stem cell-like characteristics both in vivo and in vitro. The purpose of this study was to generate and characterize novel Müller glial cell lines from the postnatal mouse retina.
METHODS: Cells were cultured from postnatal day (P) 10 double heterozygous transgenic (H-2K(b)-tsA58/+; HRhoGFP/+) or C57BL/6 mice after papain dissociation. Interferon gamma (IFNγ) induction of the SV40 T-antigen (TAg) was assayed by immunohistochemistry and Western blot analysis. Proliferation was assayed by BrdU uptake and cell counts of calcein AM/ethidium bromide-stained cells. Gene expression was analyzed by RT-PCR and immunohistochemistry.
RESULTS: Conditionally immortalized (ImM10 [Immortmouse Müller P10]) and spontaneously immortalized (C57M10 [C57BL/6 Müller P10]) Müller glial cell lines were selected by differential adherence to laminin; both consisted of adherent flat cells with large, diffusely staining nuclei and an epithelial morphology. TAg induction stimulated BrdU uptake by Müller glia in mixed retinal cultures from H-2K(b)-tsA58/+; HRhoGFP/+ mice and increased the proliferation of ImM10 cells. ImM10 and C57M10 cells expressed genes characteristic of Müller glia but not genes characteristic of differentiated retinal neurons. ImM10 cells also expressed retinal stem cell genes.
CONCLUSIONS: The ImM10 cell line is a novel, conditionally immortalized Müller glial cell line isolated from the P10 mouse retina that expresses genes characteristic of Müller glial and retinal stem cells.
Authors:
Deborah C Otteson; M Joseph Phillips
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-05-26
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  51     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-10-28     Completed Date:  2010-12-07     Revised Date:  2013-05-29    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5991-6000     Citation Subset:  IM    
Affiliation:
Department of Vision Science, College of Optometry, University of Houston, Houston, Texas 77204-2020, USA. dotteson@optometry.uh.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Animals, Newborn
Antigens, Polyomavirus Transforming / metabolism
Biological Markers / metabolism
Blotting, Western
Cell Count
Cell Line
Cell Proliferation
Cell Separation
DNA Replication
Eye Proteins / genetics*,  metabolism
Female
Fluorescent Antibody Technique, Indirect
Gene Expression / physiology*
Male
Mice
Mice, Inbred C57BL
Mice, Transgenic
Neuroglia / cytology*,  metabolism
Retinal Neurons / cytology*,  metabolism
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells / cytology*,  metabolism
Transcription Factors / genetics*,  metabolism
Grant Support
ID/Acronym/Agency:
NEIT32 EY07024/EY/NEI NIH HHS; PN30 EY07751/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, Polyomavirus Transforming; 0/Biological Markers; 0/Eye Proteins; 0/Transcription Factors
Comments/Corrections

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