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ID:32120516
大小:8.37 MB
页数:62页
时间:2019-01-31
《低氧下共培养诱导黄韧带干细胞向髓核样细胞分化的实验分析》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、万方数据第三军医大学硕+学位论文<0.05).3.ARerl4days’cocultllreunderhypoxia,real一timePCRandwestem-blottingresultsrevealedthatcomparedwithLFSCsgrownundernonnoxia,hypoxia—treatedLFSCsexpressedhigherleVelsofSox一9,coUagen—II,aggrecan,KRT19,CAl2,FOXFl,andHIF—l0【genesandHIF一10【,
2、collagen—II,Sox一9,andaggrecanproteins(P<0.05).4.LFSCsdif氨:rentiationpathwaywasidentifiedbycellsurfacemarl(ers,andourdatasuggestthatthesurfacemarkerexpressionwouldchangedduringdifferentiation.Atthefirstday,CDl05andCD271highlyexpress.GD2,Tie2andCD24haVelow1e
3、Vel.At14days,A1mostaUofthesemarkersdown—regulateexceptforCD24,andCD24expressup—regulation.Affer2ldays,onlyCD24andCD271werepositiVe,butotherswerenegatiVe.Immunocytochemistrystainingofspecialproteinsshowsthatcollagen—IIandaggrecanexpressat14and2ldays.Theexpr
4、essionofc01lagen—Idecreasesfollowingthecellsdiljf÷rentiation.5.0VerexpressionofHIF-1旺resultedinincreaseastrongalcianbluestainingofcartilagematrixandmatIlrechondrocyteswasobservedinsidethechondrogenicnodulesat3and7days,comparedtoGFP一1entirVimsinfectedcontro
5、ls.Realtime·PCRshowsthatHIF一1aoverexpressionenhanceofchondrogenesisactsViadown—regulationofthetranscriptionfactorTwist-1,andalsoenhancetheexpressionofconagen-IIandaggrecan(P<0.05).AquickresponseofTwist—ltoHIF—la,ImmunonuorescenceusingTwist一1antibodyinLFSCs
6、showedthatTwist一1wasdecreased48ha盘eroVer—expressionofHIF-lQ.QuantificationofTwist一1proteinleVelinWestemblotswasperformedusingImageJsoRwareshowsthatdecreaseofTwistlwithover-expressionofHIF一1aat6,24,48h.Conchlsions:1.Thisstudypresentsthepresenceofstemcellsin
7、thehumanLF,whichobtainedbytransforaminalendoscopicsu唱eryormicroendoscopicdiscectomysu唱ery.Wefoundthat8pg/cm2Fibronectinand4U/m1thrombin—mediatedadhesionhassuperiority.Wealsoidenti母surfacemarkerStro一1wasnegatiVe.Interestingly,ourdatashowthatstemcellmarkersO
8、ct4aIllpli6cationsweresigni6cantlydetectedinLFSCs.Oct4isrequiredf.oremcientself-renewalandthepluripotencyofstemcells,andourresultssuggestthatLFSCswassignificantlyexpressedcomparedtothatofB】ⅥSCstomaintainpluri
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