脂肪源性干細(xì)胞體外成骨特性的研究

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1、脂肪源性干細(xì)胞體外成骨特性的研究作者：李俊剛，王萬明，孫效棠【摘要】　　目的探討兔脂肪源性干細(xì)胞體外成骨分化能力。方法取3個月齡新西蘭白兔頸背部皮下脂肪組織，Ⅰ型膠原酶消化獲得細(xì)胞。繪制細(xì)胞生長曲線，采用免疫細(xì)胞熒光法檢測細(xì)胞表面抗原CD44、CD45；分別加入條件培養(yǎng)液對所獲得細(xì)胞進(jìn)行成骨成脂誘導(dǎo)，并分別進(jìn)行形態(tài)學(xué)、堿性磷酸酶、鈣鹽沉積和油紅O染色相關(guān)檢測；將脂肪源性干細(xì)胞分為成骨誘導(dǎo)組和非成骨誘導(dǎo)組，分別于誘導(dǎo)后1、2、3、4周檢測兩組的堿性磷酸酶和鈣離子濃度并作統(tǒng)計分析。結(jié)果a)所獲細(xì)胞CD44陽性表達(dá)，CD45陰性表達(dá)；所繪制的生長曲線成典型的“

2、S”型。b)在誘導(dǎo)條件下，堿性磷酸酶及鈣鹽沉積均呈陽性表達(dá)，油紅O染色為陽性。c)堿性磷酸酶濃度在誘導(dǎo)組與非誘導(dǎo)組之間存在組間差別，在各個時間點誘導(dǎo)組堿性磷酸酶濃度均高于非誘導(dǎo)組(P＜，n＝5)；誘導(dǎo)組堿性磷酸酶濃度在不同誘導(dǎo)時相其濃度變化趨勢不同(P＜，n＝5)；鈣離子濃度在誘導(dǎo)組與非誘導(dǎo)組之間存在組間差別，在各個時間點誘導(dǎo)組鈣離子濃度高于非誘導(dǎo)組(P＜，n＝5)。誘導(dǎo)組鈣離子濃度在不同誘導(dǎo)時相其濃度變化的趨勢不同(P＜，n＝5)。結(jié)論脂肪干細(xì)胞的易分離培養(yǎng)、增殖快和良好的成骨誘導(dǎo)活性完全符合對種子細(xì)胞的要求，是一種理想的骨組織工程種子細(xì)胞。【關(guān)鍵詞】

3、脂肪源性干細(xì)胞；成骨分化；堿性磷酸酶；鈣離子　　Abstract：ObjectiveToexplorethedifferentiationofadiposederivedstemcells(ADSCs)fromrabbitsintoosteogeniccellsinInthisstudy，ADSCsweregainedbycollagenaseⅠdigestionofsubcutaneousfatfromthenapeofNewZealandwhitesurfaceantigenCD44、CD4wereidentifiedbyimmunofluore

4、scenceandcellauxodromewereADSCswereperformedosteogenicandadipogenicdifferentiationrespectively，followingwithtextsrelatedmorphology，alkalinephosphataseassay，VonKossaandrathonumredOweredividedintoosteogenicinducedgroupandosteogenicnoninducedphosphataseassayandcalciumconcentration

5、ofthesetwogroupswerecheckedandstatisticsfromthecheckwereanalyzedat1，2，3，4wafterthesetwogroupswerebothThisstudygotfollowingresults：a)ADSCsCD4expressionwerepositiveandADSCsCD4expressionnegative；cellauxodromeswereS)Alkalinephosphataseassay，VonKossaandrathonumredOstainofADSCswereallp

6、ositiveundertheinfluenceofosteogenicandadipogenicdifferentiation)Alkalinephosphataseassays(ALP)weredifferentofthesetwogroupsALPofinducedgroupswashigherthanthatofnoninducedgroupateachdifferenttime(P＜，n＝5)andALPoftheinducedgroupwasdistinctindifferenttendencyunderdifferentinduct

7、ion(P＜，n＝5).Calciumconcentrationweredifferentofthesetwogroupscalciumconcentrationofinducedgroupswashigherthanthatofnoninducedgroupateachdifferenttime(P＜，n＝5)andcalciumconcentrationoftheinducedgroupwasdistinctindifferenttendencyunderdifferentinduction(P＜，n＝5).ConclusionThisstud

8、ysuggeststhatADSCscanbeisolatedandcultur