短期体外传代培养对兔骨髓间充质干细胞生长及成软骨能力影响.pdf

欠豆期F卜夕卜下专甲亡龙合习农对兔骨耸旬旬J七皿冷干细fi包5匕谁完L3A义车欠骨能 -少的翻多响 中岁K当断 巷 【结论】1.应用贴壁分离法可以获得形态较为一致的兔骨髓MSCs，并通过体 外培养扩增至较多的细胞量。 2.所得到的兔骨髓MSCs的生长能力强，生长曲线呈+S型，且各 代具有一致的生长趋势。短期内体外传代培养对兔骨髓MSCs的 生长能力影响不大。 3.成功地体外诱导单层兔骨髓 MSCs向软骨方向分化，软骨特异的 细胞外基质增多，特异性的Col11表达明显增多。 4短期传代的各代兔骨髓MSCs都能向软骨方向分化，具有相近的 成软骨能力。短期体外传代培养对兔骨髓MSCs的成软骨能力没 有明显的影响。 5.骨髓MSCs可通过体外短期传代培养，从而在数量上达到软骨组 织工程应用的需要，而且其成软骨能力仍能保持。 【关键词】骨髓间充质干细胞;传代培养;成软骨分化;转化生长因子一Rl, 兔 X豆期k卜夕卜下专十吃乡合刁仁R寸多匕阅,幻旬93匕质月卜全田月包月匕主-SA眨车丸1胃,A巨二今的翻乡够1 电仁摘 要 ‘ TheInfluenceofShortTerm-PassageCultivation认vitroon theProliferationandChondrogenicDifferentiationofRabbit BoneMarrowDerivedMesenchymalStemCells Abstract [Objective] Toobservethedifferenceoftheabilityofproliferationand chondrogenicdifferentiationoftherabbitbonemarrowderivedmesenchymalstem cells(MSCs)duringtheinvitropassagecultivationinshortterm [Methods] MSCswereobtainedfrom about3m1bonemarrowwhichwas aspiratedfromadultNewZealandRabbit,andwerepurifiedwithadheringand mediumchanging.L-DMED+20%FBSwasusedasculturemedium.MSCsobtained wereculturedinvitroandpassagedseveraltimes.Thechangeofmorphologywas observedduringthepassagecultivationofMSCs.Immunohistochemistrywasused todetecttheexpressionofCD71.Thecellgrowthcurvesof2nd,3`d,4thpassageof MSCsweredescribedandcompared.Eachofthesethreepassagewasdividedinto twogroups:onewasinducedgroup,whichwasculturedunderthechondrogenic differentiationcondition(l0%FBS+H-DMEM,TGF-p，l0ng/ml,Vc50gg/ml, Dex10-7mol/LInsulin51xg/ml).Theotherwascontrolgroup,whichwascultured withl0%F31S+H-DMEM.Thetoluidinebluestainwasusedduringtheinduced period.RT-PCRwasappliedtodetecttheexpressionofmRNAofColIIaftertwo weeksinducedcultivation.SPSS11.5wasusedtoanalyzeandcomparetheRTPCR results [Results] PurifiedMSCsderivedfromrabbitw