brdu标记对大鼠骨髓间充质干细胞增殖和成骨能力的影响

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1、BrdU标记对大鼠骨髓问充质干细胞增殖与成骨能力的影响福建医科大学附属厦门第一医院骨科研究生：冯进益导师：芮钢教授中文摘要目的：观察5一溴．2一脱氧尿嘧啶核苷(5．bromo．2-deoxyuridine，BrdU)标记前后对SD大鼠骨髓间充质干细胞(Mesenchymalstemcells，MSCs)增殖与成骨能力的影响。方法：直接贴壁法分离培养大鼠骨髓MSCs，0．2％Brdu标记后检测其增殖能力、克隆形成能力；经成骨诱导液诱导培养3周后，利用茜素红染色，荧光免疫组化和RealTimePCR观测标记前后MSCs的成骨能力的变

2、化情况。结果：BrdU标记后MSCs克隆形成能力明显减弱，在克隆个数和面积上均小于未标记组(尸<0．05)；在增殖能力方面，BrdU标记后第4d开始进入对数增长期时增殖较慢，其中第6d，8d，12d和14d两组之间有显著性差别，且BrdU标记组增殖高峰较低妒0．05)。结论：BrdU标记抑制MSCs的克隆增殖形成能力，但不会降低

3、其成骨分化能力，可以广泛应用于MSCs作为种子细胞的干细胞研究的示踪剂。关键词：BrdU；骨髓问充质干细胞；组织工程EffectsofproliferationandosteogenicpotentialityinratbonemarrowmesenchymalstemcellbyusingBrdUlabelingAbstractobjective：Toobservetheeffectsofproliferationandosteogenicpotentialityofratbonemarrowmesenchymalstemce

4、lls(MSCs)byusingBrdUlabeling．Methods：Ratmarrowderived．MSCswereisolatedandadherentcultured．Theabilityofmonocloneandproliferationweredetectedafterlabeling晰th0．2％BrdU．MeanwmletheMSCswereinducedtoosteoblastbyosteogenicliquidfor3weeks．Osteogenicpotentialitywasobservedbyal

5、izarinbordeaux，fluorescenceimmunohistochemistryandRealTimePCR．Results：TheproliferationandmonocloneofMSCsweredecreasedobviouslyafterBrdUlabeling．Thenumberandareaofclonesinlabledgroupwerelessthantheywereinunlabeledgroup伊

6、fthelabelinggroupWassignificantlylowerthanitwasinunlabledin6d，8d，12dand14d，andthepeakwaslowerthantheunlabledone．Astheosteogenicpotentiality,twogroupscellswereexpressedosteocalcinandcollagenandwereappearencedcalciumnodules．However,theRT-PCRreportedthattherewasnosignif

7、icantdifferenceintheexpressionofosteocalcingenebetweenthetwogroups(p>O．05)．Conclusions：TheproliferationandmonoclonewereinhibitedbylabeledBrdU，buttheabilityoftheosteogenicwasnotbedecreased．Therefore，BrdUlabelingCanbe一2-appliedwidelyonthefieldofMSCsasseedcellsfortheinv

8、estigationofstemcells．KeywordsBrdU；Mesenchymalstemcells；Tissueengineering．3．MSCsMesenchymalstemcellsBrdU5-bromo--2--deoxyuridine英语缩