全骨髓法体外培养分离大鼠骨髓间充质干细胞及pkh26 标记

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1、www.CRTER.org叶小凤，等.全骨髓法体外培养分离大鼠骨髓间充质干细胞及PKH26标记*全骨髓法体外培养分离大鼠骨髓间充质干细胞及PKH26标记**★叶小凤，何援利，王雪峰，付霞霏BonemarrowmesenchymalstemcellsisolatedfromratsbywholebonemarrowadherentcultureinvitroandPKH26labelingYeXiao-feng,HeYuan-Li,WangXue-feng,FuXia-feiDepartmentofObstetricsandGynecology,ZhujiangH

2、ospital,SouthernMedicalUniversity,Guangzhou510282,GuangdongProvince,ChinaYeXiao-feng★,Studyingformaster’sdegree,DepartmentofObstetricsandGynecology,ZhujiangHospital,SouthernMedicalUniversity,Guangzhou510282,GuangdongProvince,China320367@163.comCorrespondenceto:HeYuan-li,Chiefphysicia

3、n,Professor,DepartmentofObstetricsandGynecology,ZhujiangHospital,SouthernMedicalUniversity,Guangzhou510282,GuangdongProvince,ChinaSupportedby:theNationalNaturalScienceFoundationofChina,No.81041101*;theNaturalScienceFoundationofGuangdongProvince,No.10451051501004704*Received:2010-12-0

4、3Accepted:2011-01-17AbstractBACKGROUND:Invitroculturing,amplificationandlabelingareimportantlinkstoharvestlabeledratbonemarrowmesenchymalstemcells(BMSCs)thataresatisfactorytoanimalexperiment.OBJECTIVE:ToexploreaconvenientandpracticalmethodforseparatingandlabelingBMSCsusingwholebonema

5、rrowadherentculture,andlabelingwithPKH26invitro.METHODS:BMSCswereisolatedandcultivatedfromthebonemarrowofratsbywholebonemarrowadherentculture.Furtherpurificationwasachievedbyexpansionatserialpassages.Thepassage3BMSCswereculturedandlabeledwithPKH26.Thegrowth,fluorescenceintensityandse

6、rialsubcuhivationoflabeledBMSCswereanalyzedwithfluorescencemicroscope.TheproliferationabilityoftheselabeledcellswastestedbyMTT.RESULTSANDCONCLUSION:BMSCswereisolatedandpurifiedsuccessfullyandeffectivelybythemethodofwholebonemarrowadherentculture.ThelabeledBMSCsappearedredfluorescence

7、.After3passagesofserialsubcultivation,thefluorescenceintensityandthelabelingrateofBMSCsweregraduallydecreased.Thebiologicalfeaturessuchasmorphology,growthinvitrowerenotaffectedbylabeling.BMSCscanbesuccessfullycultivatedbywholebonemarrowadherencemethodconveniently.LabelingtheBMSCswith

8、PKH26isaneff