Expression of Stem Cell Markers in the Human Fetal Kidney 英文参考文献.docVIP

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Expression of Stem Cell Markers in the Human Fetal Kidney 英文参考文献.doc

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Expression of Stem Cell Markers in the Human Fetal Kidney 英文参考文献

ExpressionofStemCellMarkersintheHumanFetal Kidney SallyMetsuyanim1,OritHarari-Steinberg1,EllaBuzhor1,DoritOmer1,NaomiPode-Shakked1,HerzlBen- Hur2,ReuvitHalperin2,DavidSchneider2,BenjaminDekel1* 1DepartmentofPediatricsandPediatricStemCellResearchInstitute,ShebaMedicalCenter,TelHashomer,SacklerFacultyofMedicine,TelAvivUniversity,TelAviv,Israel, 2DepartmentofObstetricsandGynecology,AssafHarofeMedicalCenter,Zeriffin,Israel Abstract In the human fetal kidney (HFK) self-renewing stem cells residing in the metanephric mesenchyme (MM)/blastema are induced to form all cell types of the nephron till 34th week of gestation. Definition of useful markers is crucial for the identificationofHFKstemcells.Becausewilms’tumor,apediatricrenalcancer,initiatesfromretentionofrenalstemcells, wehypothesizedthatsurfaceantigenspreviouslyup-regulatedinmicroarraysofbothHFKandblastema-enrichedstem-like wilms’ tumor xenografts (NCAM, ACVRIIB, DLK1/PREF, GPR39, FZD7, FZD2, NTRK2) are likely to be relevant markers. Comprehensiveprofilingoftheseputativeandofadditionalstemcellmarkers(CD34,CD133,c-Kit,CD90,CD105,CD24 )in mid-gestationHFKwasperformedusingimmunostainingandFACSinconjunctionwithEpCAM,anepithelialsurfacemarker thatisabsentfromtheMMandincreasesalongnephrondifferentiationandhencecanbeseparatedintonegative,dimor brightfractions.NomarkerwasspecificallylocalizedtotheMM.Nevertheless,FZD7andNTRK2werepreferentiallylocalized totheMMandemergingtubules(,10%ofHFKcells)andweremostlypresentwithintheEpCAMnegandEpCAMdim fractions, indicatingputativestem/progenitormarkers.Incontrast,singlemarkerssuchasCD24andCD133aswellasdouble-positive CD24 CD133+ cells comprise .50% of HFK cells and predominantly co-express EpCAMbright, indicating they are mostly markers of differentiation. Furthermore, localization of NCAM exclusively in the MM and in its nephron progenitor derivativesbutalsoinstromaandtheexpressionpatternofsignificantlyelevatedrenalstem/progenitorgenesSix2,Wt1, Cited1,andSall1inNCAM+EpCAM-andtoalesse