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Isolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation.

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dc.contributor.author이주카요코-
dc.contributor.author홍사민-
dc.contributor.author김찬윤-
dc.contributor.author성공제-
dc.date.accessioned2014-12-19T17:28:38Z-
dc.date.available2014-12-19T17:28:38Z-
dc.date.issued2012-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/91480-
dc.description.abstractPURPOSE: To establish an effective system for isolating primary retinal ganglion cells (RGCs) from newborn mice. METHODS: The retinas were separated from enucleated eyeballs of Crl:CD-1 mice on postnatal day 1 to 4. RGCs were purified using three different methods, including two-step immunopanning (TSI), direct magnetic separation (DMS), and immunopanning-magnetic separation (IMS). Harvested cells were maintained for 24 h in a defined medium and then examined with immunocytochemistry, western immunoblotting, and real-time reverse transcription polymerase chain reaction (RT-PCR) for glial cell-specific glial fibrillary acidic protein (GFAP) and amacrine cell-specific syntaxin 1. RESULTS: As determined with immunofluorescence staining, RGCs purified by TSI were sparsely mixed with GFAP-positive astrocytes, and RGCs isolated by DMS were frequently mixed with syntaxin 1-positive amacrine cells. However, RGCs collected by IMS were seldom contaminated by GFAP-positive or syntaxin 1-positive cells. On western immunoblots, TSI cells showed significant GFAP expression, and DMS cells showed apparent syntaxin 1 expression, but IMS cells did not. Results of the real-time RT-PCR showed a similar tendency to those of the immunocytochemistry and western immunoblots. CONCLUSION: Primary mouse RGCs were highly purified by the IMS method, combining the benefits of the TSI and DMS methods. This isolation method may provide a good experimental system for studying glaucoma in vitro.-
dc.description.statementOfResponsibilityopen-
dc.relation.isPartOfMOLECULAR VISION-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.subject.MESHAmacrine Cells/cytology-
dc.subject.MESHAmacrine Cells/metabolism-
dc.subject.MESHAnimals-
dc.subject.MESHAnimals, Newborn-
dc.subject.MESHAntibodies/chemistry-
dc.subject.MESHAntibodies/immunology-
dc.subject.MESHAstrocytes/cytology-
dc.subject.MESHAstrocytes/metabolism-
dc.subject.MESHBiomarkers/metabolism-
dc.subject.MESHBlotting, Western-
dc.subject.MESHFemale-
dc.subject.MESHGene Expression-
dc.subject.MESHGlial Fibrillary Acidic Protein-
dc.subject.MESHImmunohistochemistry-
dc.subject.MESHImmunomagnetic Separation/methods*-
dc.subject.MESHImmunomagnetic Separation/standards-
dc.subject.MESHMice-
dc.subject.MESHNerve Tissue Proteins/genetics-
dc.subject.MESHNerve Tissue Proteins/metabolism-
dc.subject.MESHPrimary Cell Culture-
dc.subject.MESHReal-Time Polymerase Chain Reaction-
dc.subject.MESHRetinal Ganglion Cells/cytology*-
dc.subject.MESHRetinal Ganglion Cells/metabolism-
dc.subject.MESHSyntaxin 1/genetics-
dc.subject.MESHSyntaxin 1/metabolism-
dc.titleIsolation of primary mouse retinal ganglion cells using immunopanning-magnetic separation.-
dc.typeArticle-
dc.contributor.collegeCollege of Medicine (의과대학)-
dc.contributor.departmentDept. of Ophthalmology (안과학)-
dc.contributor.googleauthorSamin Hong-
dc.contributor.googleauthorYoko Iizuka-
dc.contributor.googleauthorChan Yun Kim-
dc.contributor.googleauthorGong Je Seong-
dc.identifier.doi23233794-
dc.admin.authorfalse-
dc.admin.mappingfalse-
dc.contributor.localIdA03162-
dc.contributor.localIdA04395-
dc.contributor.localIdA01035-
dc.contributor.localIdA01946-
dc.relation.journalcodeJ02272-
dc.identifier.eissn1090-0535-
dc.identifier.pmid23233794-
dc.subject.keywordAmacrine Cells/cytology-
dc.subject.keywordAmacrine Cells/metabolism-
dc.subject.keywordAnimals-
dc.subject.keywordAnimals, Newborn-
dc.subject.keywordAntibodies/chemistry-
dc.subject.keywordAntibodies/immunology-
dc.subject.keywordAstrocytes/cytology-
dc.subject.keywordAstrocytes/metabolism-
dc.subject.keywordBiomarkers/metabolism-
dc.subject.keywordBlotting, Western-
dc.subject.keywordFemale-
dc.subject.keywordGene Expression-
dc.subject.keywordGlial Fibrillary Acidic Protein-
dc.subject.keywordImmunohistochemistry-
dc.subject.keywordImmunomagnetic Separation/methods*-
dc.subject.keywordImmunomagnetic Separation/standards-
dc.subject.keywordMice-
dc.subject.keywordNerve Tissue Proteins/genetics-
dc.subject.keywordNerve Tissue Proteins/metabolism-
dc.subject.keywordPrimary Cell Culture-
dc.subject.keywordReal-Time Polymerase Chain Reaction-
dc.subject.keywordRetinal Ganglion Cells/cytology*-
dc.subject.keywordRetinal Ganglion Cells/metabolism-
dc.subject.keywordSyntaxin 1/genetics-
dc.subject.keywordSyntaxin 1/metabolism-
dc.contributor.alternativeNameIizuka, Yoko-
dc.contributor.alternativeNameHong, Sa Min-
dc.contributor.alternativeNameKim, Chan Yun-
dc.contributor.alternativeNameSeong, Gong Je-
dc.contributor.affiliatedAuthorIizuka, Yoko-
dc.contributor.affiliatedAuthorHong, Sa Min-
dc.contributor.affiliatedAuthorKim, Chan Yun-
dc.contributor.affiliatedAuthorSeong, Gong Je-
dc.citation.volume18-
dc.citation.startPage2922-
dc.citation.endPage2930-
dc.identifier.bibliographicCitationMOLECULAR VISION, Vol.18 : 2922-2930, 2012-
dc.identifier.rimsid31303-
dc.type.rimsART-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Ophthalmology (안과학교실) > 1. Journal Papers
1. College of Medicine (의과대학) > Yonsei Biomedical Research Center (연세의생명연구원) > 1. Journal Papers

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