notch基因在人瘢痕疙瘩间充质干细胞中的表达

邓呈亮，王达利，朱晶晶等.notch基因在人瘢痕疙瘩间充质干细胞中的表达[J].中华整形外科杂志.2014,30(3):198-202.

基因;瘫痕，肥大性;干细胞

邓呈亮，王达利，朱晶晶，王波，余丽梅

遵义医学院附院烧伤整形外科

2014年

198-202

知网

目的研究notch1-4基因在人瘢痕疙瘩间充质干细胞中的表达,探讨notch信号通路是否参与瘢痕疙瘩的形成。方法收集瘢痕疙瘩样本,二步酶消化法收获人瘢痕疙瘩间充质干细胞,用流式细胞仪分析原代和P3代细胞表型;免疫细胞化学检测Oct4、波形蛋白和CK19的表达;体外诱导瘢痕疙瘩间充质干细胞向成骨细胞分化,茜素红S检测钙盐沉积情况;通过荧光定量PCR检测瘢痕疙瘩间充质干细胞notch1-4 mRNA的 Objective To study the expressions of Notchl一gene in human keloid-derived mesenehymal-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 mesenchymal-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 cells 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-PGR. The relative quantitative of Notch2，Notch3 gene were higher than Notchl， Notch4 gene(P。