冻存对CM-Dil标记脐带间充质干细胞的影响.pdf

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1、·214·国际生物医学工程杂志2014年8月第37卷第4期IntJBiomedEng，August2014，Vol37，No．4·论著·冻存对CM—Dil标记脐带间充质干细胞的影响陈艳王建军李怀芳【摘要】目的考察用氯甲基苯甲酰氨(CM—Dil)标记人脐带间充质干细胞(hUCMSCs)的可行性及冻存对CM—Dil标记的hUCMSCs的影响。方法采用组织块法体外培养hUCMSCs，观察细胞形态及生长情况，通过流式细胞仪检测细胞免疫表型来鉴定该细胞。将第3代hUCMSCs用CM．Dil标记，观察细胞形态及生长情况

2、，用荧光显微镜观察体外标记情况，并将细胞冻存、复苏，观察细胞复苏后的生长情况及荧光标记情况。结果hUCMSCs形态类似成纤维细胞，呈梭形、多角形。第3代hUCMSCs强表达CD44、CD29、CD90和CD73，不表达CD34和CD45。CM．Dil标记hUCMSCs的细胞标记率达90％以上，染料主要存在于细胞膜；标记后细胞生长状态良好，冻存及复苏后细胞仍有较强荧光表达，且细胞生长状态良好。结论CM—Dil标记hUCMSCs示踪方法简单易行，冻存对CM—Dil标记的hUCMSCs的生长无明显影响。【关键词】

3、人脐带间充质干细胞；氯甲基苯甲酰氨；低温冻存；示踪中图分类号：R318；Q813文献标识码：A文章编号：1673—4181(2014)04—0214—04Impactofcry0preservati0nonCM-DillabeledhumanumbilicalcordmesenchymalstemcellsChenYah,WangJianjun，LiHuaifang．DepartmentofObstetriesandGynecology,AfiliatedTongfiHospitdofTon~iUniver

4、sity,Shanghai200065,China【Abstract】ObjectiveToobservethefeasibilityoflabelinghumanumbilicalcordmesenchymalstemcells(hUCMSCs)withchloromethyl—benzamidodialkylcarbocyanine(CM—Dil)andtheinfluenceofcryopreservationontheCM—DillabeledhUCMSCs．MethodsUsingtissueex

5、planttocuhurehUCMSCs，themorphologyandgrowthsituationofthecellswereobserved，andcellphenotypewereidentifiedbyflowcytometry．Theceilsofthe3rdgenerationwerelabeledwithCM—Dil．Thecellmorphology，growthsituationandfluorescentmarkerswereobservedunderfluorescentmicro

6、scope．Thelabeledcellswerecryopreservedandrecovered，andthegrowthofcellsanditsfluorescentlabelingconditionswereobservedafterrecovery．ResultsThemorphologyofhUCMSCswassimilartofibroblasts，spindle—shapedandpolygona1．Thecellsofthe3rdgenerationofhUCMSCsstronglyex

7、pressedCD44，CD29，CD90andCD73，butdidnotexpressCD34andCD45．ThemarkrateofCM—DiltolabelhUCMSCsreachedashighas90％andthedyewasmainlyinthecellmembrane．Thelabeledcellsareingoodconditionandthecellsaftercryopreservationandrecoverystillhavestrongfluorescenceexpressio

8、n，alsothecellgrowthisingoodcondition．ConclusionsUsingCM—DiltolabelandtrackhUCMSCsissimplerthanothermethods．CryopreservationdoesnothavenegativeinfluenceonCM．Dil—labeledhUCMSCs．【Keywords】Humanumbilicalcordmesen