Differential Adhesion Molecule Expression during Murine Embryonic Stem Cell Commitment to the Hematopoietic and Endothelial Lineages 英文参考文献.docVIP

DifferentialAdhesionMoleculeExpressionduring MurineEmbryonicStemCellCommitmenttothe HematopoieticandEndothelialLineages BashaL.Stankovich,EsmeraldaAguayo,FatimaBarragan,AniketSharma,MariaG.Pallavicini* SchoolofNaturalSciences,UniversityofCaliforniaMerced,Merced,California,UnitedStatesofAmerica Abstract Mouseembryonicstemcells(ESC)makecellfatedecisionsbasedonintrinsicandextrinsicfactors.ThedecisionofESCto differentiate to multiple lineages in vitro occurs during the formation of embryoid bodies (EB) andis influenced by cell- environmentinteractions.However,molecularmechanismsunderlyingcell-environmentalmodulationofESCfatedecisions areincompletelyunderstood.Sinceadhesionmolecules(AM)influenceproliferationanddifferentiationindevelopingand adult tissues, we hypothesized that specific AM interactions influence ESC commitment toward hematopoietic and endothelial lineages. Expression of AM in the adherens, tight and gap junction pathways in ESC subpopulations were quantified.E-cadherin(E-cad),Claudin-4(Cldn4),Connexin-43(Cx43),ZonaOccludens-1(ZO-1)andZonaOccludens-2(ZO- 2)transcriptlevelsweredifferentiallyexpressedduringearlystagesofhematopoietic/endothelialcommitment.StableESC linesweregeneratedwithreducedexpressionofE-cad,Cldn4,Cx43,ZO-1andZO-2usingshRNAtechnology.Functional and phenotypic consequences of modulating AM expression were assessed using hematopoietic colony forming assays, endothelial sprouting assays and surface protein expression. A decrease in E-cad, Cldn4, Cx43 and ZO-1 expression was associatedwithlesscommitmenttothehematopoieticlineageandincreasedendothelialdifferentiationasevidencedby functional and phenotypic analysis. A reduction in ZO-2 expression did not influence endothelial differentiation, but decreasedhematopoieticcommitmenttwo-fold.ThesedataindicatethatasubsetofAMinfluenceESCdecisionstocommit to endothelial and hematopoietic lineages. Furthermore, differentially expressed AM may provide novel markers to delineateearlystagesofESCcommitmentto