骨髓间充质干细胞分化的成骨细胞支持脐血造血干祖细胞的研究.doc

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1、骨髓间充质干细胞分化的成骨细胞支持脐血造血干祖细胞的研究作者：黄晓兵，刘霆，孟文彤，智伟【摘要】本研究利用骨髓间充质干细胞诱导分化的成骨细胞构建二维培养体系，探讨其体外支持脐血干/祖细胞生存的作用。体外进行人骨髓间充质干细胞（MSC）原代培养后诱导为成骨细胞，构建二维培养体系。将免疫磁珠分选的CD34+脐血干/祖细胞接种于其中，在无外源性细胞因子情况下进行体外培养，进行CFU，HPP-CFU和LTC-IC测定并将其分别与MSC、成骨细胞，MSC/成骨细胞（1∶2）作为饲养细胞的二维培养体系实验结果相比较。结果发现：在所

2、构建的造血干/祖细胞（HSPC）二维培养体系中，骨髓MSC诱导成骨细胞对于脐血造血干/祖细胞培养的支持作用显著优于其他各组培养体系，尤其对长期造血干细胞（longterm-HSC）体外生存有更为明显地支持作用。结论：骨髓间充质干细胞诱导成骨细胞构建的二维体系对于脐血干/祖细胞体外培养具有支持作用，进一步证实成骨细胞对造血干细胞生存与增殖具有重要调控作用。【关键词】骨髓间充质干细胞　　OsteoblastsDifferentiatedfromHumanMarrowBoneMesen-chymalStemCellsSupp

3、ortHematopoieticStem/ProgenitorCellsfromUmbilicalCordBlood　　AbstractThisstudywasaimedtoconstructatwo-demensionalculturesystembyusingosteoblastsinducedfromhumanmarrowmesenchymalstemcells(MSC)andtoinvestigateitssupporteffectonsurvivalofhematopoieticstem/progenitor

4、cellsforumbilicalcordblood(UCB)exvivo.MSCswereisolatedfromadulthumanbonemarrowandwerecultured，thesecondgenerationofMSCswereinducedintoosteoblastswhichwereirradiatedwith20GygammaraysinaCobalt60sourceandconfluencedintoafeederlayer.CD34+cellswereselectedfromfreshum

5、bilicalcordbloodsamplesbyusingMicrobeadKitofMiniMACSandseededintothetwo-dimensionalculturesystemtocultureexvivowithoutexogenouscytokines.Byusingcolony-formingassay，highproliferativepotentialcolony-formingcellassay，andlong-termcultureinitiatingcellassay，theabilit

6、yofthetwo-dimensionalsystemtocultureHSCs/HPCswasobserved.TheresultsshowedthattheosteoblastsinducedfrombonemarrowMSCinconstructedtwo-demensionalculturesystemdisplayedmoresignificantsupporteffectonsurvivalofhematopoieticstem/progenitorcellsfromumbilicalcordblood(U

7、CB)exvivo，comparedwithotherculturesystems，especiallyonlongtermHSCs7survivalexvivo.Itisconcludedthatthetwo-dimensionalculturesystemconstitutedbyosteoblastsinducedfromhumanMSCshascertainabilityofsupportingmaintenanceandmultipotencyofHSCs/HPCsfromumbilicalcordblood

8、invitro，especiallysustainingsurvivalofHSCinlong-termculture.IthasalsobeenprovedthatosteoblastsplayacrucialroleinregulationofHSCgrowth.　Keywordsmarrowmesenchymalstemce