MYC Gene Delivery to Adult Mouse Utricles Stimulates Proliferation of Postmitotic Supporting Cells In Vitro 英文参考文献.docVIP

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MYC Gene Delivery to Adult Mouse Utricles Stimulates Proliferation of Postmitotic Supporting Cells In Vitro 英文参考文献.doc

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MYC Gene Delivery to Adult Mouse Utricles Stimulates Proliferation of Postmitotic Supporting Cells In Vitro 英文参考文献

MYCGeneDeliverytoAdultMouseUtriclesStimulates ProliferationofPostmitoticSupportingCellsInVitro JosephC.Burns*,JamesJ.Yoo,AnthonyAtala,JohnD.Jackson WakeForestInstituteforRegenerativeMedicine,WakeForestSchoolofMedicine,Winston-Salem,NorthCarolina,UnitedStatesofAmerica Abstract Theinnerearsofadulthumansandothermammalspossessalimitedcapacityforregeneratingsensoryhaircells,whichcan leadtopermanentauditoryandvestibulardeficits.Duringdevelopmentandregeneration,undifferentiatedsupportingcells withininnerearsensoryepitheliacanself-renewandgiverisetonewhaircells;however,theseoticprogenitorsbecome depletedpostnatally.Therefore,reprogrammingdifferentiatedsupportingcellsintooticprogenitorsisapotentialstrategy forrestoringregenerativepotentialtotheear.Transientexpressionoftheinducedpluripotencytranscriptionfactors,Oct3/ 4,Klf4,Sox2,andc-Mycreprogramsfibroblastsintoneuralprogenitorsunderneural-promotingcultureconditions,soasa firststep,weexploredwhetherectopicexpressionofthesefactorscanreversesupportingcellquiescenceinwholeorgan culturesofadultmouseutricles.Co-infectionofutricleswithadenoviralvectorsseparatelyencodingOct3/4,Klf4,Sox2,and thedegradation-resistantT58Amutantofc-Myc(c-MycT58A)triggeredsignificantlevelsofsupportingcellS-phaseentryas assessed by continuous BrdU labeling. Of the four factors, c-MycT58A alone was both necessary and sufficient for the proliferativeresponse.ThenumberofBrdU-labeledcellsplateauedbetween5–7daysafterinfection,andthendecreased ,60% by 3weeks, as many cycling cells appeared to enter apoptosis. Switching to differentiation-promoting culture medium at 5days after ectopic expression of c-MycT58A temporarily attenuated the loss of BrdU-labeled cells and accompaniedaverymodestbutsignificantexpansionofthesensoryepithelium.Asmallnumberoftheproliferatingcellsin thesecultureslabeledforthehaircellmarker,myosinVIIA,suggestingtheyhadbegundifferentiatingtowardsahaircell fate.Theresultsindicatethatectopicexpressionofc-MycT58Aincombinationwithmethodsforpromo