大鼠来源滑膜间充质干细胞的成软骨分化邵博，龚忠诚，刘慧，凌彬，克热木•阿巴斯，尹小朋，胡露露，王冰，宁晓婷，杨萌，林兆全新疆医科大学第一附属医院颌面肿瘤外科，新疆医科大学口腔医学院，新疆维吾尔自治区口腔医学研究所，新疆维吾尔自治区乌鲁木齐市830054Chondrogenicdifferentiationofratsynovial-derivedmesenchymalstemcellsShaoBo,GongZhong-cheng,LiuHui,LingBin,KeremuAbass,YinXiao-peng,HuLu-lu,WangBing,NingXiao-ting,YangMeng,LinZhao-quanDepartmentofOral&MaxillofacialOncologySurgery,theFirstAffiliatedHospitalofXinjiangMedicalUniversity,StomatologyCollegeofXinjiangMedicalUniversity,StomatologyResearchInstituteofXinjiangUygurAutonomousRegion,Urumqi830054,XinjiangUygurAutonomousRegion,China背景：相比其他组织来源的间充质干细胞，滑膜间充质干细胞有着较强的成软骨特性和克隆能力，因此将是软骨组织工程中最有前景的种子细胞之一。目的：培养及体外扩增SD大鼠滑膜间充质干细胞，鉴定其多向分化潜能及在添加生长因子后三维立体培养条件下的成软骨能力。方法：无菌条件下获取SD大鼠滑膜组织，Ⅰ型胶原酶消化法分离大鼠滑膜间充质干细胞。体外扩增、培养，取第3代滑膜间充质干细胞进行吉姆萨、生长曲线测定、成脂诱导、成骨诱导和三维条件下成软骨诱导，成软骨诱导21d后通过甲苯胺蓝染色、Ⅱ型胶原免疫组化染色及RT-PCR对成软骨诱导后产物进行检测。结果与结论：获取的大鼠滑膜细胞具有间充质干细胞的特性，滑膜间充质干细胞在体外培养呈成纤维样形态，并维持着多向分化的能力。使用生长因子诱导软骨细胞21d后，可见软骨样组织，蛋白聚糖和Ⅱ型胶原可以在甲苯胺蓝染色和Ⅱ型胶原免疫组化染色后被检测到。RT-PCR检测结果显示软骨诱导分化后细胞中Ⅱ型胶原和蛋白聚糖mRNA呈阳性表达。提示大鼠来源的滑膜间充质干细胞具有成软骨能力。中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程全文链接：关键词：组织构建；软骨组织工程；干细胞；分化；滑膜间充质干细胞；成骨；成脂；成软骨；生长因子；三维培养；国家自然科学基金；Abstract：BACKGROUND:Comparedwithothersourcesofmesenchymalstemcells,synovial-derivedmesenchymalstemcellshavesignificantcharacteristicsofchondrogenesisandcloning.Therefore,synovial-derivedmesenchymalstemcellsareoneofthemostpromisingseedcellsincartilagetissueengineering.OBJECTIVE:Toisolateandculturesynovial-derivedmesenchymalstemcellsofSprague-Dawleyrats,identifythemultipotentialdifferentiationandthepotentialabilityofchondrogenicdifferentiationinthree-dimensionalculturecondition.METHODS:ThesynoviumtissuewasharvestedfromSprague-Dawleyrats.Thesynovial-derivedmesenchymalstemcellswereisolatedwithtypeⅠcollagenenzymedigestionmethodandculturedinvitro.Thepassage3cellsweredetectedwithgiemsastaining,thecellcycle,adipogenicandosteogenicdifferentiationweredetermined.Thepassage3cellswerecentrifugedaspelletsandculturedinthechondriogenicmediumfor21days.Andthepelletswereexaminedbytoluidinebluestaining,typeⅡcollagenimmunohistochemicalstainingandRT-PCR.RESULTSANDCONCLUSION:Themesenchymalstemcellsisolatedfromthesynoviumtissueofratshavethecharacteristicsofmesenchymalstemcells,andexhibitfibroblast-likemorphologyafterculturedinvitro.Themultilineagedifferentiationpotentialswerealsorevealed.Afterthecellwereculturedinchondrogenicmediumfor21days,chondroidtissuewasfound,typeIIcollagenandaggrecancouldbedetectedpositivelybytoluidinebluestaining,typeⅡcollagenimmunohistochemicalstaining,andexpressedbyRT-PCRexamination.Therefore,synovialmesenchymalstemcellshaveachondrogenicdifferentiationpotential.中国组织工程研究杂志出版内容重点：...