兔脂肪干细胞体外成骨诱导培养及成骨活性鉴定的实验研究张开刚1张月东1王玫2（1.泰安市中心医院骨科，山东泰安2710002.泰山医学院，山东泰安271016）【摘要】目的体外分离培养兔脂肪干细胞，在成骨诱导条件下，鉴定其成骨活性。方法取4月龄新西兰大白兔腹股沟区脂肪,Ⅰ型胶原酶消化,分离、培养及传代原细胞,将第3代ADSCs消化后，加入成骨培养液中诱导分化,分别行碱性磷酸酶染色、茜素红染色、VonKossa（钙结节）染色鉴定分化结果.结果体外分离培养的细胞增殖活跃，成骨诱导后检测碱性磷酸酶染色、茜素红染色、VonKossa（钙结节）染色均呈阳性表达。结论脂肪干细胞具有成骨分化能力,是一种理想的骨组织工程种子细胞。【关键词】脂肪干细胞；成骨诱导；种子细胞Anexperimentalstudyofosteogeniccultureinvitroandidentificatingtheosteogenicactivityofrabbitadipose-derivedstemcellsZHANGKaigang1,ZHANGYuedong1,WangMei2（1.DepartmentofOrthopedicSurgery,TaianCenterHospital,ShandongTaian,2710002.TaishanMedicalCollege，ShandongTaian271016）【Abstract】ObjectiveSeparatingandCulturingrabbitadipose-derivedstemcells，identificatingtheosteogenicactivityinosteogenicconditions.MethodsExtractinginguinalfatofNewZealandwhiterabbitsaged4months,UsingIcollagenasedigestiong,separating,culturingandpassagingoftheoriginalcells,addingosteogenicmediumtoinduceosteogenicdifferentiationafterdigestingthe3rdgenerationADSCs,thenidentificatingthedifferentiationresultsbyalkalinephosphatasestaining、alizarinredstaining、VonKossa(calciumnodules)staining.ResultsThecellsofSeparatingandCulturinginvitrowereactive,thealkalinephosphatasestaining、alizarinredstainingandVonKossa(calciumnodules)stainingwerebothpositiveexpressionafterosteogenic.ConclusionAdiposederivedstemcellshavetheabilityofOsteogenicdifferentiation,theyareanidealseedcellsforbonetissueengineering.【Keywords】Adipose-DerivedStemCells;Osteogenic;SeedCellsAnExperimentalstudyofAdiposestemcellsculturedinvitroosteogenicandosteogenicactivityidentificationZHANGKaigang1,ZHANGYuedong1,WangMei2（1.DepartmentofOrthopedicSurgery,TaianCenterHospital,ShandongTaian,2710002.TaishanMedicalCollege，ShandongTaian271016）【Abstract】ObjectiveIsolationandCultureofadiposestemcellsinosteogenicconditionstoidentifyosteogenicactivity.MethodsExtractinginguinalfatofNewZealandwhiterabbitsaged4months,UsingIcollagenasedigestion,separation,cultureandpassageoftheoriginalcells,the3rdgenerationADSCsafterdigestion,byaddingosteogenicmediumtoinduceosteogenicdifferentiation,alkalinephosphatasestainingwereperformed,alizarinredstaining,VonKossa(calciumnodules)stainingdifferentiationresults.ResultsProliferationofisolatedandculturedcellsculturedinvitroActiveisactive,Detectionofbonealkalinephosphatasestainingafterinduction,alizarinredstaining,VonKossa(calciumnodules)werebothpositiveforexpression.ConclusionAdiposederivedstemcellshavetheabilitytodifferentiateintobone,itisanidealseedcellsforbonetissueengineering.【Keywords】Adipose-DerivedStemCells;Osteogenic;SeedCells脂肪干细胞（adipose-derivedstemcells，ADSCs）是近年来发现的一种成体干细胞，2001年，Zuk〔1〕等首次从人的脂肪组织悬液中分离出了具有与骨髓间充质干细胞类似，可以在体外稳定增殖、具有多向分化潜能的脂肪干细胞。因具有来源丰富、取材方便、对机体的损伤小、增殖快、能多向分化等优点,迅速成为组织工程学、再生医学等领域的研究热点。本实验通过分离培养兔脂肪干细胞，研究其成骨分化能力，探讨其作为骨组织工程种子细胞的可行性。1．材料和方法1.1材料1.1.1主要试剂和仪器I型胶原酶（sigma美国）；胰蛋白酶（sigma美国）；地塞...
