中文摘要：

目的 研究Notch基因在人瘢痕疙瘩间充质样干细胞中的表达,探讨Notch信号通路是否参与瘢痕疙瘩的形成.方法 收集6例瘢痕疙瘩患者的瘢痕疙瘩样本,二步酶消化法获取人瘢痕疙瘩间充质样干细胞,流式细胞仪分析原代和第3代细胞表型;免疫细胞化学检测细胞胚胎蛋白4（Oct4）、波形蛋白和CK19的表达;体外诱导细胞向成骨细胞分化,茜素红S检测钙盐沉积情况;实时定量PCR检测细胞Notch1-4 mRNA的表达.结果 流式细胞术结果显示原代及第3代瘢痕疙瘩间充质样干细胞均高表达间充质干细胞表型CD29 （94.93％）、CD44（89.79％）、CD90 （74.28％）,不表达造血干细胞表型CD34（1.26％）、CD45 （0.79％）;免疫细胞化学结果显示表达多能干细胞标志Oct4和间充质细胞标志波形蛋白,不表达上皮细胞标志CK19;向成骨细胞体外诱导21d后,可见明显钙盐结节;Realtime-PCR结果显示Notch1-4基因在瘢痕疙瘩间充质样干细胞中均有表达,且Notch2、Notch3基因的相对表达量高于Notch1、Notch4基因（P＜0.05）.结论 人瘢痕疙瘩间充质样干细胞Notch基因不同亚型的表达差异可能与其自我更新、增殖、分化及参与瘢痕疙瘩形成有关.

英文摘要：

Objective To study the expressions of Notchl-4 gene in human keloid-derived mesenchymal-like stem cells, and to explore the Notch signaling pathway＇ s role in the formation of keloid. Methods Keloid samples were collected to harvest human keloid-derived mesenchymal-like stem cells through two-step enzymatic dissociation method. By flow cytometry, cell phenotype of primary and P3 generation were analyzed. By immunocytochemistry, the expressions of Oct4, vimentin and CK19 were examined. Keloid-derived mesenehymal-like stem cells were induced into osteoblasts in vitro and calcium deposition was detected by Alizarin red S stain. Realtime polymerase chain reaction（RT-PCR） was used to detect the expressions of Notchl-4 mRNA in keloid-derived mesenchymal-like stem cells. Results Flow cytometry showed that keloid-derived mesenchymal-like stem ceils of primary and P3 generation highly expressed CD29, CD44, CD90 from the typical MSC phenotype marker, but they failed to express HSC phenotype markers, such as CD34 and CD45. The results of immunocytochemistry showed that Oct4 from pluripotent stem cell markers and vimentin from mesenchymal cell markers was positive and CK19 from epithelial cell markers was negative. After induced differentiation into osteoblasts in vitro after 21 day, calcium nodules could be seen clearly; Notchl-4 gene were expressed in keloid-derived mesenchymal-like stem cells through RT-PCR. The relative quantitative of Notch2, Notch3 gene were higher than Notehl, Notch4 gene （P 〈 0.05）. Conclusions The expression difference of different subtypes from Notch gene in human keloid-derived mesenchymal-like stem cells may be related to self-renewal, proliferation, differentiation, and participate in the formation of keloid.