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ID:20226248
大小:60.00 KB
页数:5页
时间:2018-10-08
《基质细胞衍生因子》由会员上传分享,免费在线阅读,更多相关内容在学术论文-天天文库。
1、基质细胞衍生因子.freelalCell-derivedFactor1andPlateletFactor4ontheAdhesionCharacteristicsandChemotacticFunctionofExVivoExpandedUmbilicalCordBloodCD34+CellsAbstractToinvestigatetheeffectsofstromalcell-derivedfactor1(SDF-1)andplateletfactor4(PF4)onthehoming-relatedfunct
2、ionofexpandedexvivoumbilicalcordbloodCD34+cells,purifiedcordbloodCD34+cells-freemediumcontainingaHGFbinationofFL+SCF+TPO(FST)lSDF-1alone,100ng/mlPF4alone,orbothofthese2cytokines.TheexpansionrateofCD34+cells,colonyformation,homing-relatedfunctionsincludingexpre
3、ssionofhoming-relatedadhesionmoleculesofexpandedCD34+cell,adhesionactivityandchemotacticfunctionofthere-selectedexpandedCD34+cellsepoints.TheresultsshoultipleofCFUinSDF-1groupsarkableup-regulated,incontrast,expressionofCXCR-4ontheexpandedCD34+cellsarkabledoark
4、ablyup-regulatedinthePF4groups.InalltheSDF-1group,PF4groupandSDF-1plusPF4group,theabilityofexpandedCD34+cellsadheringtofibronectinlayerigrationrateofexpandedCD34+cellsinSDF-1groupsigrationrateigrationrateinPF4groupsigrationrateofexpandedCD34+cellsintheSDF-1plu
5、sPF4groupsoleculesonexpandedCD34+cells,andretaintheadherentandmigrationabilityofexpandedCD34+cells,ingofexpandedCD34+cells.Inshort,SDF-1andPF4arehelpfulforthehoming-relatedfunctionoftheexpandedUCBHSPC.Keybilicalcordblod;stromalcell-derivedfactor1;plateletfacto
6、r4;adhesioncharacteristics;chemotacticfunction;ex-vivoexpansion理想的体外扩增方案应该在扩增HSPC的同时保持其增殖、自我更新及向骨髓(BM)微环境归巢功能的完整。我们曾报道过短期体外扩增并不影响HSPC的归巢相关功能,但随着扩增时间延长和部分归巢相关黏附分子的表达,黏附功能和迁移功能呈下降趋势[1]。在此基础上我们进一步研究了在扩增体系中加入基质细胞衍生因子1(SDF-1)和PF4对脐血(umbilicalcordblood,UCB)CD34+细胞的归
7、巢相关功能的影响,探讨通过优化UCBHSPC的体外扩增因子组合方案,在扩增UCBHSPC的同时保持其归巢相关功能。材料和方法标本来源和细胞制备无菌采集健康足月产新生儿脐血,应用Ficoll-Hypaque淋巴细胞分离液(比重1.077)分离出单个核细胞(MNC)。按照MACSCD34+细胞单选试剂盒(MiltenyiBiotchInc.)说明书分离纯化CD34+细胞。体外扩增培养将纯化的CD34+细胞根据试验目的按2×104/ml接种于5-15mlStemlineTMII无血清培养液(Sigma)中,加入早期作用因
8、子FL+SCF+TPO(FST),其中含有50ng/ml的SCF、100ng/ml的TPO和FL,并于接种0天添加1次IL-320ng/ml,将培养瓶置于37℃、5%的CO2孵箱中培养14天。培养第4天加入等体积新鲜培养液(含相同浓度的TPO和FL,半量浓度的SCF)。培养第7天、第10天变体积换液,维持换液后细胞浓度为2×105/ml左右,并添加与第4天相
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