Differing Lectin Binding Profiles among Human Embryonic Stem Cells and Derivatives Aid in the Isolation of Neural Progenitor Cells 英文参考文献.docVIP

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Differing Lectin Binding Profiles among Human Embryonic Stem Cells and Derivatives Aid in the Isolation of Neural Progenitor Cells 英文参考文献.doc

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Differing Lectin Binding Profiles among Human Embryonic Stem Cells and Derivatives Aid in the Isolation of Neural Progenitor Cells 英文参考文献

DifferingLectinBindingProfilesamongHuman EmbryonicStemCellsandDerivativesAidintheIsolation ofNeuralProgenitorCells MaheshC.Dodla1,AmberYoung1,AlisonVenable2,KowserHasneen1,RajR.Rao3,DavidW.Machacek4, StevenL.Stice1,4 * 1RegenerativeBioscienceCenter,UniversityofGeorgia,Athens,Georgia,UnitedStatesofAmerica,2DepartmentofBiochemistryandMolecularBiology,Universityof Georgia, Athens, Georgia, United States ofAmerica, 3Department ofChemicaland Life Sciences Engineering, Virginia Commonwealth University, Richmond, Virginia, UnitedStatesofAmerica,4ArunABiomedicalInc.,Athens,Georgia,UnitedStatesofAmerica Abstract Humanembryonicstemcells(hESCs)andtheirdifferentiatedprogenyallowforinvestigationofimportantchanges/events duringnormalembryonicdevelopment.Currentlymostoftheresearchisfocusedonproteinacouschangesoccurringasa resultofdifferentiationofstemcellsandlittleisknownaboutchangesincellsurfaceglycosylationpatterns.Identificationof cell lineage specific glycans can help in understanding their role in maintenance, proliferation and differentiation. Furthermore,theseglycanscanserveasmarkersforisolationofhomogenouspopulationsofcells.Usingapanelofeight biotinylatedlectins,theglycanexpressionofhESCs,hESCs-derivedhumanneuralprogenitors(hNP)cells,andhESCs-derived mesenchymalprogenitor(hMP)cellswasinvestigated.OurgoalwastoidentifyglycansthatareuniqueforhNPcellsand use the corresponding lectins for cell isolation. Flow cytometry and immunocytochemistry were used to determine expressionandlocalizationofglycans,respectively,ineachcelltype.Theseresultsshowthattheglycanexpressionchanges upondifferentiationofhESCsandisdifferentforneuralandmesenchymallineage.Forexample,bindingofPHA-Llectinis lowinhESCs(1464.4%)butsignificantlyhigherindifferentiatedhNPcells(9960.4%)andhMPcells(9063%).Threelectins: VVA,DBAandLTLhavelowbindinginhESCsandhMPcells,butsignificantlyhigherbindinginhNPcells.Finally,VVAlectin bindingwasusedtoisolatehNPcellsfromamixedpopulationofhESCs,hNPcellsandhMPcells.Thisisthefirstr