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1、中国实验血液学杂志JournalofExperimentalHematology2004;12(2):133-137·133·文章编号(ArticleID):1009-2137(2004)02-0133-05·论著·无血清脐血巨核系祖细胞体外扩增的研究莫文健,毛平,何秋山,应逸,朱志刚,许艳丽,杜庆华广州医学院附属广州市第一人民医院血液内科,广州510180摘要 脐血造血干细胞移植后血小板恢复延迟是一大难题,目前认为这主要与脐血中巨核系祖细胞数量不足及脐血巨核细胞分化成熟延迟有关,而将部分脐血进行巨核系祖细胞体外扩增后输注受者
2、体内是有望解决这一难题的重要途径,但适用于临床应用的扩增条件至今仍未确立。本课题采用人脐血单个核细胞(MNC)在无血清培养体+系中使用TPO,IL23,SCF,IL26等细胞因子进行不同的组合,在培养的0,6,10,14天进行MNC、CD41细胞及CFU2MK数的检测,以寻找最佳的细胞因子组合及最佳的收获时机。结果表明:无血清条件下TPO与IL23,SCF,IL26等细胞因子联用可实现脐血巨核系祖细胞有效的体外扩增,各因子组中以TPO+IL23+SCF+IL26组扩增效+果为最佳,其CFU2MK数于第10天最多,扩增达6.8倍,
3、CD41细胞扩增达8.8倍。结论:在人脐血MNC无血清培养条件下,TPO+IL23+SCF+IL26组为巨核系祖细胞体外扩增较佳的因子组合。由于TPO+IL23+SCF+IL26+组的CFU2MK数于第10天最多,CD41细胞数亦为同期最高,故培养后收获时间宜控制在其体外培养的第10天。关键词 脐血;巨核系祖细胞;无血清培养;血小板;巨核细胞+中图分类号 R329.28文献标识码 AExvivoExpansionofMegakaryocyteProgenitorCellsfromHumanCordBloodinSerum2
4、freeCultureMOWen2Jian,MAOPing,HEQiu2Shan,YINGYi,ZHUZhi2Gang,XIUYan2Li,DUQing2HuaDepartmentofHematology,InternalMedicineofAffiliatedGuangzhouFirstMunicipalPeople′sHospital,GuangzhouMedicalCollege,Guangzhou510180,ChinaAbstractProlongedthrombocytopeniaisapuzzlingproblem
5、followingumbilicalcordblood(CB)transplantation.Itmightbetheresultofinadequatemegakaryocyteprogenitorsandthearrestinthemegakaryocytematuration.Itisanimportantmethodtosolvetheproblembytransfusingex2vivoexpandedCBmegakaryocyteprogenitorcellsintothepatientstoshortenthedu
6、rationofplateletrecovery.However,themostoptimalconditionofexpansionhasnotbeenestablishedsofar.Inthestudy,cordbloodmononuclearcells(MNC)wereculturedinserum2freemediumwithTPO,+IL23,SCFandIL26.ThenumbersofMNC,CFU2MKandCD41cellsweremeasuredat0,6,10and14days,inordertofind
7、thebestcytokinecombinationandoptimalharvesttimepointforclinicaluse.TheresultsshowedthatthemegakaryocyteprogenitorsmostefficientlyexpandedwiththecytokinecombinationincludingTPO,IL23,SCFandIL26.+ThetimepointofmaximalCFU2MKgrowthisday10.At10days,thenumbersofCFU2MKandCD4
8、1cellsexpandedby6.82and8.82foldrespectively.Inconclusion,invitro,thecytokinecocktailincludingTPO,IL23,SCFandIL26wasthemostoptimalcy
