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1、【摘要】目的探讨不同大鼠脂肪基质细胞的培养、增殖与纯化方法，初步研究其生物学特性。方法取大鼠皮下和肠系膜部位的脂肪组织，根据两种组织对胶原酶的敏感性不同，获取基质细胞，观察生长情况，并进行鉴定。结果不同品系大鼠脂肪含量不同，皮下和肠系膜脂肪基质干细胞产率不同，生物学特性相似，贴畢细胞分两类：第一类有明显汕滴；第二类细胞内无明显汕滴。第一类细胞经过传代，逐渐消失，第二类细胞所要获取的脂肪基质细胞，组化鉴定CD49d(+)和CD106G),为脂肪基质细胞表型。结论建立了一种大鼠脂肪基质细胞的培养、增殖与纯化方法，不同部位来源的脂肪组织酶消化时间和细胞产率不同，
2、同时发现所获得细胞分两类，性质冇所不同，为脂肪基质细胞的相关研究打下一定的细胞生物学基础。【关键词】大鼠脂肪肪基质细胞培养Isolation,CultureandIdentificationofRatAdipose?DeriveclStemCellsTnVitroCHENLian?feng,LINXue,FANGQuan(DepartmentofCardiology,PUMCIbChineseAcademyofMedicalSciencesandPekingUnionMedicalCollege,Beijing,China)【Abstract]object
3、iveToexplorethemethodologyregardingculture,proliferationandpurificationofadipose?derivedstemcelIs(ADSCs),andtostudytheirbiologicalcharacteristics.MethodsFattissuewasobtainedfrommesenteryandsubclermaltissuesofratsofdifferentlineages・Thetissuesamplesweredigestedwithcollagenasetoobta
4、inadiposestromalcells・Cellswereisolatedandpurifiedbasedontheirdifferentreactiontodigestion.Immunohistochoniistrywasusedtocharacteri^eandidentifytheharvestedcells.Growthofcellswasobservedwith■aninvertedcontrastphasemicroscope.ResultsDespitedifferentproductivity,adiposestromalcellsf
5、romdifferentsourcetissuesanddifferentlineagesshowedidenticalbiologicalcharacteristics・After2?3daysofculture,floatingcellsbecameadherentandgrewintoclusters.Afterseveralpassages,thesecelIsseparatedandchangedintospindle?shaped・TheadherentcelIscouldbefurtherclassifiedintotwotypes.Thef
6、irsttypeoflargersizecontaineclprofuselipiddropletsandhighlypositivebyoilred0staining.Thesecondtypewasofslimmer?shape,withlittleintracellularlipidandnegativeoilred0staining・Afterpassages,remainingfloatingcells,albeitinasmal1number,continueslydifferentiatingintocellsofthesecondtype,
7、whilecellsoffirsttypegraduallydiminished.Assessedbyimmunohistochemistry,cellsofthesecondtypewereCD49d(+)andCD106(-),whichwerecharacteristicphenotypeofadiposestromalcells.ConclusionWithmodifieddurationofdigestion,ourmethodcanbeusedtoculture,proliferate,andpurifyadiposestromalcells,
8、despitediffereneeregardingproduct
