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1、滑膜干细胞向成骨细胞及软骨细胞分化的实验研究#**51015(广州中医药大学第一附属医院三骨科,广州,510405)摘要:目的:探讨滑膜间充质干细胞(SMSCs)向成骨细胞及软骨细胞分化特性。方法:将第四代SMSCs以5×106接种于3块6孔板,其中一块板诱导SMSCs向软骨细胞分化,剩下2块板诱导SMSCs向成骨细胞分化,每板4孔,2孔为空白对照孔,另2孔为观察组,2天后换液,加入诱导液。空白组加入含10%胎牛血清(FBS)的L-DMEM培养基继续培养,成骨诱导组在基础培养基中上加入成骨诱导剂(含β甘油磷酸钠10mmo
2、l/L、地塞米松0.1umol/L、维生素C50mg/L),软骨细胞组为在基础培养基中加入软骨诱导剂(含转化生长因子β110μg/L、地塞米松0.1μmol/L、维生素C50mg/L。在第14天取出1块6孔板做阿尔新蓝染色,1块6孔板做碱性磷酸酶染色;第20天取出剩余6孔板做茜素红染色。结果:SMSCs经软骨诱导液培养后,细胞体积增大,由梭形变为椭圆形状,部分细胞核偏位,呈肾形,培养14后可见细胞周围有基质分泌,阿尔新蓝染色阳性。SMSCs经成骨诱导液培养后细胞从梭形变为类圆形,细胞体积明显增加,14天时细胞集落的表面可见有
3、颗粒状物质分泌,碱性磷酸酶染色为阳性,21天时茜素红染色阳性,表现出成骨细胞的特点。结论:SMSCs广泛存在于滑膜组织内,表现出干细胞特性,在特定条件下可向成骨细胞及软骨细胞分化。关键词:滑膜间充质干细胞;成骨细胞;软骨细胞中图分类号:R-33120ExperimentalstudyonSynovialmesenchymalstemcellsdifferentiationintoosteoblastsandchondrocytesZengJianchun,FanYueguang,ZengYirong,LiJie,LiFeil
4、ong(Thethirddepartmentoforthopaedics,ThefirstaffiliatedhospitalofGuangzhouuniversityof2530354045Chinesemedicine,Guangzhou,510405)Abstract:Objective:Toexplorefeaturesofsynovialmesenchymalstemcells(SMSCs)differentiationintotheosteoblastandchondrocytes.Methods:Thefou
5、rthgenerationofSMSCswereseededin36-wellplatesby5×106,inoneoftheplatesSMSCswereinducedtochondrocytes,intheremainingtwoboardSMSCswereinducedtodifferentiateintoosteoblast,4holedineachplate,twoholeswereblankholes,andtheothertwoholesweretheobservationgroup,twodayslatert
6、heculturemediumwaschangedwithinducingfluid.ThecontrolgroupwasaddedL-DMEMmediumcontaining10%fetalbovineserum(FBS),theinductinggroupwasaddedinducingagentonthebasisoftheL-DMEMmedium(osteogenicgroupcontainingβglycerophosphate10mmol/L,dexamethasone0.1umol/L,vitaminC50mg
7、/L;thechondrogenicgroupcontainintransforminggrowthfactorβ110μg/L,dexamethasone0.1μmol/L,vitaminC50mg/L,insulin6.25mg/L,transferrin6.25ug/ml,bovineserumalbumin1.25mg/ml.Onthe14thdayremoveda6-welldoneAlcianbluestaining,a6-wellplatedonealkalinephosphatasestaining;onth
8、e21stdayremovetheremaining6-wellplatedonealizarinredstaining.Results:VolumeofSMSCsincreasedafterinducedbychondroinductivemedium,thespindlechanged