Protective Effect of Human Amniotic Fluid Stem Cells in an Immunodeficient Mouse Model of Acute Tubular Necrosis 英文参考文献.docVIP

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Protective Effect of Human Amniotic Fluid Stem Cells in an Immunodeficient Mouse Model of Acute Tubular Necrosis 英文参考文献.doc

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Protective Effect of Human Amniotic Fluid Stem Cells in an Immunodeficient Mouse Model of Acute Tubular Necrosis 英文参考文献

AmnioticStemCellsandKidney Results induction as compared with the control that did not undergo glycerol induced muscle damage (Figure 2C). The difference in thenumberofapoptoticcellspresentintheglyceroltreatedmice when compared with the untreated control mice was highly statisticallysignificant(P,0.01)(Figure2E). hAFSCPhenotypeandKaryotypebeforeInjection hAFSC,beforeinjection,presentafibroblastoidshapeasshown in Figure 1A. The cells were then tested to confirm a normal karyotype before in vivo applications, to exclude chromosomal abnormalitiesthatcouldcompromisetheirpluripotentialcapabil- ity(Figure1B).hAFSCwereanalyzedfortheexpressionofearly andlatekidneymarkersbeforeinjection.AsshowninFigure1C, prior to injection, hAFSC were negative for the most important kidney markers, ranging from transcription factors expressed during early kidney development to late differentiation markers. Thus,weconfirmedthathAFSCarenotspecificallycommittedto kidneyprogenitorcellswhenculturedinvitro. InVivoDetectionofhAFSCbyBioluminescence hAFSC, transduced with a lentivirus coding for luciferase, showed stable expression of the transgene over many population doublings.Thecellsnotinfectedwiththelentivirusandexposedto luciferin (the luciferase substate) did not reveal any biolumines- cence,whilethetransfectedcellsexposedtoluciferincontinueto express the bioluminescent signal after 20 population doublings (Figure3A). InFigure3Bisshownaninvitroexperimenttodeterminethe minimal number (16105) of cells that exhibited a readily detectable opticalsignal.InFigure3Cisshownliveimagingof a right kidney injected with 1,26106 hAFSC after damage EvaluationoftheGlycerolInducedMuscleDamageand ATNUsingPeriodAcidSchiffStaining(PAS)andTUNEL Staining Figure2Ademonstratesnormalmorphologyofanu/numouse kidney before any damage; the proximal and distal tubules are intactaswellastheglomeruli.Figure2Bshowsmorphologyof thekidney3daysafterintramuscularinjectionofglycerol.Marked disorganization of the normal st