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

《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