Sorting Live Stem Cells Based on Sox2 mRNA Expression 英文参考文献.docVIP

SortingLiveStemCellsBasedonSox2mRNAExpression HansM.Larsson1,SeungTaeLee1,2,MartaRoccio3,DianaVelluto1,4,MatthiasP.Lutolf3,PeterFrey1,5, JeffreyA.Hubbell1* 1Laboratory for Regenerative Medicine and Pharmacobiology, Institute for Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fe′de′raledeLausanne,Lausanne,Switzerland,2LaboratoryofStemCellBiomodulation,DepartmentofAnimalBiotechnology,KangwonNationalUniversity,Chuncheon, Korea,3LaboratoryofStemCellBioengineering,InstituteforBioengineering,SchoolofLifeSciences,EcolePolytechniqueFe′de′raledeLausanne,Lausanne,Switzerland, 4SchoolofBiologicalandChemicalSciences,QueenMary,UniversityofLondon,London,UnitedKingdom,5LaboratoryofExperimentalPediatricUrology,Departmentof PediatricUrology,CentreHospitalierUniversitaireVaudois,Lausanne,Switzerland Abstract While cell sorting usually relies on cell-surface protein markers, molecular beacons (MBs) offer the potential to sort cells basedonthepresenceofanyexpressedmRNAandinprinciplecouldbeextremelyusefultosortrarecellpopulationsfrom primary isolates. We show here how stem cells can be purified from mixed cell populations by sorting based on MBs. Specifically,wedesignedmolecularbeaconstargetingSox2,awell-knownstemcellmarkerformurineembryonic(mES)and neural stem cells (NSC). One of our designed molecular beacons displayed an increase in fluorescence compared to a nonspecificmolecularbeaconbothinvitroandinvivowhentestedinmESandNSCs.WesortedSox2-MB SSEA1+cellsfrom a mixed population of 4-day retinoic acid-treated mES cells and effectively isolated live undifferentiated stem cells. Additionally, Sox2-MB+ cells isolated from primary mouse brains were sorted and generated neurospheres with higher efficiency than Sox2-MB2 cells. These results demonstrate the utility of MBs for stem cell sorting in an mRNA-specific manner. + Citation:LarssonHM,LeeST,RoccioM,VellutoD,LutolfMP,etal.(2012)SortingLiveStemCellsBasedonSox2mRNAExpression.PLoSONE7(11):e4