诱导脐血内皮祖细胞分化成内皮细胞的研究  

Study on Inducing of Cord Blood Derived Endothelial Progenitor Cells into Endothelial Cells

作　　者：王秀华[1] 贾兵[1] 陈张根[1] 王克强[2] 

机构地区：[1]复旦大学附属儿科医院心血管中心,上海200032 [2]复旦大学附属中山医院心血管病研究所,上海200032

出　　处：《实用儿科临床杂志》2006年第20期1407-1408,1433,共3页Journal of Applied Clinical Pediatrics

摘　　要：目的探讨诱导脐血来源内皮祖细胞分化为内皮细胞,分析其用作组织工程心血管修复物种子细胞来源的可行性。方法采用6%羟乙基淀粉沉降法和密度梯度离心法联合应用分离脐血单个核细胞,在含有血管内皮生长因子(VEGF)等多种生长因子的培养液中培养扩增,通过形态学(光镜、电镜)、免疫荧光和流式细胞仪等技术对脐血来源内皮祖细胞分化的内皮细胞进行鉴定。结果从新鲜脐血中得到的单个核细胞数(3.4±2.1)×107/mL。贴壁细胞培养分化过程中形态发生改变,从小圆形变成梭形,最后分化成典型成熟内皮细胞的鹅卵石样形态,细胞数量可扩增达107。7 d后免疫荧光检测发现细胞表达内皮细胞特异性标志vWF和VEGFR-2。流式细胞仪分析单个核细胞经诱导分化后细胞表面表达CD133下降,从3.11%±1.05%下降至0.09%±0.02%。透射电镜观察到培养14 d的细胞胞浆中已出现典型的Weibel-Palade小体。结论脐血单个核细胞存在内皮祖细胞,并在体外可诱导分化为内皮细胞,初步具有用作构建组织工程心血管修复物种子细胞的可行性。Objective To induce cord blood derived endothelial progenitor cells（EPCs）into endothelial cells, and investigate the leasi bility of these cells as the seed cells of tissue engineering cardiovascular replacement. Methods Mononuclear cells were isolated from fresh cord bood hy 6%HES and density gradient centrifugation. Isolated cells were cultured in medium supplemented with vascular endothelial grows factor（VEGF）. Attached cells were identied by morphology, immunofluorescence staining and flow cytometry. Results The per centage of mononuclear cells isolated from fresh cord blood was （3.4±2.1 ）×10^7/mL. The morphology of attached cells changed while being cultured and inducted, from small sized round cells to spindle like cells, to a typical cobblestone morphology,and the total number of cells was 10^7. After 7 days of culture, immunofluorescence staining showed that the vWF and VEGFR 2 were expressed. Compared with the original, cell markers CD133 decreased（3.11%±1.05 % ） to （0.09 %±0.02 % ）, P〈0.05. After 14 days of culture, Weibel Palade bodies were shown on the transmission electron microscopy photomicrographs. Conclusions There are endothelial progenitor cells in the cord blood mononuclear cells, and they can be inducted into endothelial cells and have the feasibility of as the seed cells of tissue engineering cardiovascular replacement.

关 键 词：脐血 内皮祖细胞 内皮细胞 组织工程 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]