brdu標(biāo)記對(duì)大鼠骨髓間充質(zhì)干細(xì)胞增殖和成骨能力的影響

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1、BrdU標(biāo)記對(duì)大鼠骨髓問充質(zhì)干細(xì)胞增殖與成骨能力的影響福建醫(yī)科大學(xué)附屬?gòu)B門第一醫(yī)院骨科研究生：馮進(jìn)益導(dǎo)師：芮鋼教授中文摘要目的：觀察5一溴．2一脫氧尿嘧啶核苷(5．bromo．2-deoxyuridine，BrdU)標(biāo)記前后對(duì)SD大鼠骨髓間充質(zhì)干細(xì)胞(Mesenchymalstemcells，MSCs)增殖與成骨能力的影響。方法：直接貼壁法分離培養(yǎng)大鼠骨髓MSCs，0．2％Brdu標(biāo)記后檢測(cè)其增殖能力、克隆形成能力；經(jīng)成骨誘導(dǎo)液誘導(dǎo)培養(yǎng)3周后，利用茜素紅染色，熒光免疫組化和RealTimePCR觀測(cè)標(biāo)記前后MSCs的成骨能力的變

2、化情況。結(jié)果：BrdU標(biāo)記后MSCs克隆形成能力明顯減弱，在克隆個(gè)數(shù)和面積上均小于未標(biāo)記組(尸<0．05)；在增殖能力方面，BrdU標(biāo)記后第4d開始進(jìn)入對(duì)數(shù)增長(zhǎng)期時(shí)增殖較慢，其中第6d，8d，12d和14d兩組之間有顯著性差別，且BrdU標(biāo)記組增殖高峰較低妒0．05)。結(jié)論：BrdU標(biāo)記抑制MSCs的克隆增殖形成能力，但不會(huì)降低

3、其成骨分化能力，可以廣泛應(yīng)用于MSCs作為種子細(xì)胞的干細(xì)胞研究的示蹤劑。關(guān)鍵詞：BrdU；骨髓問充質(zhì)干細(xì)胞；組織工程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英語(yǔ)縮