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1、自组装多肽水凝胶支架中MSCs的三维培养及成骨分化周庆翰1,2，王玉明1，田志杰1，林娟1（610083成都，成都医学院生物医学系1;610041成都，西南民族大学化学与环境保护工程学院2）[摘要]目的使用一种新型人工设计自组装多肽（RADA16）水凝胶作为三维培养支架评价MSCs成骨分化情况。方法将人骨髓MSCs培养增殖后接种到水凝胶中，在成骨分化培养液中进一步培养1~3周。荧光染色法观察细胞形态和存活情况；组织学染色检测MSCsALP活性；半定量RT-PCR分析成骨特异性基因的表达。结果绝大多数MSCs在水凝胶支架内能够存活，呈纺锤样形态。诱导培养后蛋白和基因表达水平均检测到ALP活
2、性，在14天时达到峰值。骨晚期分化特异性基因BSP也有表达，且表达量随培养时间延长而增多。结论自组装多肽水凝胶为MSCs的黏附生长及向成骨细胞分化提供良好的三维微环境，有望成为极具吸引力的骨组织工程支架材料。[关键词]间充质干细胞；成骨分化；三维培养；自组装多肽[中图法分类号]R318；Q813[文献标志码]AThree-DimensionalCultureandOsteogenicDifferentiationofMSCsinSelf-AssemblingPeptideHydrogelScaffoldsZHOUQinghan1,2,WANGYuming1,TIANZhijie1,LIN
3、Juan1（1TheSchoolofBiomedicalScience,ChengduMedicalCollege,Chengdu,610083;2CollegeofChemistryandEnvironmentProtectionEngineering,SouthwestUniversityforNationalities，Chengdu,610041,China）[基金项目]四川省教育厅科研项目（10ZC091）；成都医学院自然科学基金资助项目（CYZ11-010）[通信作者]林娟，电话：（028）68289239，E-mail：linjuan.scu@gmail.com[Abstr
4、act]ObjectiveToevaluatetheosteogenicdifferentiationofMSCsusinganewclassofsyntheticself-assemblingpeptidehydrogels,RADA16,asascaffoldforthree-dimensionalculture.MethodsMSCsderivedfromhumanbonemarrowwereculture-expandedandseededintothehydrogelandfurtherculturedinosteogenicmediumfor1-3weeks.Thecellv
5、iabilitywasvisualizedanddetectedunderfluorescencemicroscopy.Alkalinephospharaseactivitywasevaluatedbyhistochemicaltechniques.Semi-quantitativeRT-PCRanalyseswerepreformedtodetecttheexpressionofmRNAtranscripts.ResultsAlmostMSCswerelivingcellsandhada9typicalspindleshape.Inosteogenicdifferentiation,t
6、heexpressionofALPattheproteinandmRNAlevelwassignificantlyhigherthanthatofcontrolcells.AlthoughtheALPactivitypeakedat14days,theBSPcontentincreasedcontinuouslyfor21days.ConclusionTheseresultsshowthatthebiodegradable/biocompatiblehydrogel,RADA16,supportsMSCsattachmentanddifferentiationtowardtheosteo
7、geniclineage,indicatingthatRADA16canbeconsideredasanattractivesyntheticbiomaterialforuseinbonetissueengineering.[Keywords]mesenchymalstemcells;osteogenicdifferentiation;three-dimensionalculture;self-assemblingpeptideSu
