摘要
目的 :研究来自胎儿血管壁内的CD10 5+ 细胞是否具有间充质干细胞的特性 ,地塞米松是否促进其向脂肪细胞分化。方法 :免疫组化检测CD10 5阳性的细胞在胎儿主动脉分布 ,分离主动脉血管的成纤维样细胞 ,流式细胞仪检测细胞免疫表型 ,并接种在脂肪分化和成骨分化的诱导培养液中培养 ,油红O和VonKossa染色及透射电镜鉴定脂滴和钙化基质沉淀的形成。结果 :CD10 5阳性的细胞分布在主动脉血管内膜的内皮细胞 ,部分中膜和外膜。分离到的细胞CD10 5、CD10 6、CD2 9、CD4 4阳性 ,CD34、CD31、CD11a、CD11b、HLA -DR为阴性。油红O和VonKossa染色分别显示脂滴和钙化基质形成。电镜下诱导的脂肪细胞胞浆中可见有包膜脂滴 ,诱导的成骨细胞胞浆内外电子密度高的钙盐沉积。诱导液中无地塞米松 ,未见含大脂滴的脂肪细胞。结论 :分离到主动脉壁CD10 5+ 细胞具有间充质干细胞的特性 ,并且地塞米松促进其向脂肪细胞分化 。
AIM: To investigate whether aorta-derived CD_(105)^+ cells show characteristics of mesenchymal stem cells, and if dexamethason enhances this kind of CD_(105)^+ cells to differentiate into adipocytes. METHODS: The distribution of CD105 in aorta was assessed by immunohistochemistry. The aorta wall cells were isolated and immunophenotypes were identified by FACS. CD_(105)^+ cells were sorted using MACS CD105 micromagnetic beads. The differentiation of CD_(105)^+ cells into adipocytes and osteoblasts was induced under different conditions and indicated by staining of Oil red O, detecting of alkaline phosphatase activity, calcium accumulation stained with silver nitrate and transmission electron microscope analysis, respectively. RESULTS: The endothelial cells, a part of medial smooth muscle cells and adventital fibroblasts were CD105 positive. The isolated aortic arch cells were positive for CD105, CD106, CD44, CD29, and negative for CD45, CD11a, CD11b and HLADR. The CD_(105)^+ cells differentiated into adipocytes contained Oil-Red-O-positive lipid droplets, the osteocytes with calcium deposition and alkaline phosphatase activity. Ultrastructurally, it was observed that some needle-shaped crystal calcium deposition similar to bone spicules was inside the cytoplasm of induced osteocytes. When the dexamethason was absent in the adipogenic medium, there were no adipocytes with lipid droplets. CONCLUSION: The results demonstrate that CD_(105)^+ cells show characters of MSCs reside in aortic wall, and is able to differentiate into adipocytes and osteocytes in vitro. Dexamethasone enhances aorta-derived CD_(105)^+ with characters of MSCs to differentiate into adipocytes. These suggeste that MSCs might be related with atherosclerosis. [
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2004年第10期1786-1789,共4页
Chinese Journal of Pathophysiology
基金
国家高新技术研究发展计划资助项目 ("86 3"计划 ) (No .2 0 0 2AA2 0 5 0 6 1)
ChinaMedicalBoardofNewYork Inc (Grant # 0 1- 74 8)资助