brdu, cfse和gfp標(biāo)記大鼠間充質(zhì)干細(xì)胞的比較

《brdu, cfse和gfp標(biāo)記大鼠間充質(zhì)干細(xì)胞的比較》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在學(xué)術(shù)論文-天天文庫(kù)。

1、BrdU,CFSE和GFP標(biāo)記大鼠間充質(zhì)干細(xì)胞的比較作者：付霞霏，何援利，楊芳，彭冬先，劉木彪【摘要】目的:研究BrdU,CFSE和GFP作為大鼠間充質(zhì)干細(xì)胞標(biāo)記物的可行性.方法:分別用上述三種標(biāo)記物標(biāo)記大鼠間充質(zhì)干細(xì)胞，檢測(cè)即時(shí)、15和30d的標(biāo)記率，通過(guò)檢測(cè)細(xì)胞周期、凋亡率和描繪生長(zhǎng)曲線，來(lái)明確標(biāo)記物對(duì)體外培養(yǎng)大鼠間充質(zhì)干細(xì)胞生長(zhǎng)特性的影響；通過(guò)流式細(xì)胞儀檢測(cè)標(biāo)記物對(duì)細(xì)胞表面標(biāo)志（CD29,CD34,CD44,CD45）表達(dá)的影響.結(jié)果:BrdU,CFSE,GFP的即時(shí)標(biāo)記率為77.0%,99.4%,89.7%，1mo后分別下降至51.9%,77

2、.5%,82.9%.三者對(duì)體外培養(yǎng)的大鼠間充質(zhì)干細(xì)胞的生長(zhǎng)特性和表面標(biāo)志的表達(dá)均無(wú)明顯影響.結(jié)論:GFP,BrdU和CFSE均可以作為大鼠間充質(zhì)干細(xì)胞的標(biāo)記物.【關(guān)鍵詞】綠色熒光蛋白；溴脫氧尿嘧啶核苷；熒光染料CFSE；間充質(zhì)干細(xì)胞　　【Abstract】AIM:ToinvestigatethefeasibilityoflabelingofBrdU,CFSEandGFPaslabelsofratmesenchymalstemcells(MSCs)andtoinvestigatetheinfluenceofthe3markersonthegrowthc

3、haracteristicsandexpressionsofsurfacemarkers11ofratMSCs.METHODS:WistarratsbonemarrowderivedMSCswereisolatedandpurifiedinvitrobyPercolldensitygradientcentrifugationcombinedwithadherentmethodandwerelabeledwithBrdU,CFSEandGFP,respectively.Theimmediate,15dayand30daylabelingeffic

4、iencyofthe3markerswasquantified.ThelabelingefficiencyofBrdUwasdetectedwithimmunocytochemicalmethod,andthatofCFSEandGFPwasdetectedviaflowcytometry(FCM).ThelabeledMSCswereobservedundermicroscope.CellcycleandapoptosisrateoflabeledMSCswererevealedandthegrowthcurvesweredelineated,s

5、oastoidentifywhetherornotthemarkerscastedaninfluenceonthecellgrowthcharacteristicsinvitro.Furthermore,thesurfacemarkers(CD29,CD34,CD44,CD45)ofthelabeledMSCsweredetectedthroughflowcytometryandwerecomparedwiththoseofnonlabeledMSCs.RESULTS:TheimmediatelabelingefficiencyofBrdU,CFS

6、E,andGFPwas77.0%,99.4%,89.7%respectively,andtheefficiencydecreasedto51.9%,77.5%,and82.9%respectivelyafter30d.TherewasnostatisticaldifferenceincellcycleandapoptosisratiobetweenlabeledMSCsandnonlabeledMSCs.ThegrowthcurvesoflabeledMSCsweresimilartothatofnonlabeledMSCs.The3marker

7、sdidntinfluencetheexpressionsofsurfacemarkers(CD29,CD34,CD44,CD45)ofrat11MSCs.CONCLUSION:The3markershavenoeffectsonthegrowthcharacteristicsandexpressionsofsurfacemarkersofratMSCsculturedinvitro.BrdU,CFSEandGFPcanserveaslabelingmarkersofratMSCs.BrdUandCFSElabelingcanbeusedforsh

8、orttimetracing,andGFPlabelingforlongtimetracing.　　【Key