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体外共培养人牙髓干细胞对人牙囊干细胞增殖、成骨分化的作用

, PP. 2267-2272

Keywords: 人牙囊干细胞,人牙髓干细胞,体外共培养,成骨分化

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Abstract:

目的通过体外建立人牙囊干细胞(humandentalfolliclestemcells,HDFSCs)和人牙髓干细胞(humandentalpulpstemcells,HDPSCs)共培养体系,研究HDPSCs对HDFSCs增殖、成骨分化的作用。方法①利用改良的组织块-消化联合法、免疫组化方法、流式细胞术以及多向诱导方法对HDFSCs和HDPSCs进行分离、培养、鉴定;②采用0.4μm孔径的Transwell小室构建HDFSCs和HDPSCs体外共培养体系;③CCK-8测定HDFSCs增殖活性的变化;④与HDPSCs体外共培养1周,qRT-PCR检测HDFSCs几个标志性成骨基因:碱性磷酸酶(alkalinephosphatase,ALP)、人类相关转录因子2(runt-relatedtranscriptionfactor2,Runx2)、骨桥蛋白(osteopontin,OPN)、骨涎蛋白(bonesialoprotein,BSP)以及Ⅰ型胶原(collagentype1,Col-1)的表达情况。结果①成功分离、培养、鉴定HDFSCs和HDPSCs;②从第4天开始,共培养组HDFSCs的增殖活力明显高于对照组(P<0.05);③与对照组相比,共培养组ALP、Runx2、OPN、BSP、Col-1的相对表达量均明显增高(P<0.05)。结论与HDPSCs共培养能增强HDFSCs增殖活力及成骨分化。

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