摘要
目的探讨小鼠骨髓源性血管内皮祖细胞（EPCs）体外形成管腔样结构和侵袭H22肝癌细胞的可能性和条件。方法密度梯度离心法获取小鼠骨髓中单个核细胞，利用序列差速贴壁方法（依次在培养1h和24h后提取上清液中细胞）诱导培养，噻唑蓝（MTT）比色法检测在1、3、7、10、13、16、20d的细胞增殖能力，从细胞形态、免疫荧光、细胞免疫表型对培养细胞进行鉴定并观察培养1周后细胞的体外形成管腔结构及侵袭肝癌细胞的能力。结果EPCs在1周内呈集落样生长，1周后“鹅卵石”样结构；能吞噬低密度脂蛋白和膜连接荆豆凝集素，双染阳性率（96．46±1．74）％；EPCs表达CD34（91．23±3．76）％、血管内皮生长因子受体（VEGFR）-2（92．85-±2．12）％，但CDl33（61．54±13．71）％；EPCs在能够形成管腔样结构（26．6±13．4）／高倍镜（×100）；并向肿瘤细胞富集。结论小鼠骨髓中富含EPCs，利用序列差速贴壁方法得到较高纯度EPCs，并表现成血管倾向、侵袭肿瘤组织的能力。
Objective To investigate the possibilities and terms of endothelial progenitor cells （EPCs） derived from mouse bone marrow forming the tube and invading the hepatocellular carcinoma cells H22 Clusters. Methods Bone marrow mononuelear cells were gained by density-gradient centrifugation. After culture for 1 h and 24 h, the nonadherent cells were extracted with the method of sequence of differentia] adhesion and then were induced by EGM-2 medium. Cell multiplication was tested by methyl thiazol tetrazolium （MTY） chromatometry at 1,3, 7, 10, 13, 16, 20 days. The cells were identified by cell morphology, immunofluorescence and flow cytometry analysis, and the abilities of forming the tube and invading the hepatoeellular carcinoma cells were observed after culture for 1 week. Results The EPCs grew as colony-forming units within 1 week and as cobblestones after 1 week. The cells could phagocytize the lowdensity lipoprotein and membrane binded Ulex Europaeus agglutinin with double-positive percentage （96. 46± 1.74）%. The positive expression rate of CD34 and vascular endothelial growth factor receptor （VEGFR）-2 was （91.23 ±3.76）% and （92. 85 ±2. 12）% respectively, and that of CD133 was （61.54 ± 13.71 ） %. EPCs could form tubes and recruit to the tumor cells. Conclusion EPCs were rich in mouse marrow and could be obtained by means of sequence of differential adhesion. The EPCs can form tubes and invade the malignant cell cluster.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2011年第8期1292-1294,F0003,共4页
Chinese Journal of Experimental Surgery
基金
基金项目：国家自然科学基金资助项目（30972904）
江苏省自然科学基金资助项目（BK2008063）
关键词
癌
肝细胞
内皮祖细胞
内皮细胞
成管能力
侵袭
Carcinoma, hepatocellular
Endothelial progenitor cell
Endothelial cell
Tube formation
Invasion
