中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2008年
1期
45-50
,共6页
张华蓉%陈飞兰%徐承平%卞修武
張華蓉%陳飛蘭%徐承平%卞脩武
장화용%진비란%서승평%변수무
干细胞%神经胶质瘤%新生血管化,病理性%肿瘤移植%移植,异种
榦細胞%神經膠質瘤%新生血管化,病理性%腫瘤移植%移植,異種
간세포%신경효질류%신생혈관화,병이성%종류이식%이식,이충
Stem cells%Glioma%Neovascularization,pathologic%Neoplasm transplantation%Transplantation,heterologous
目的 观察人脐血源性内皮祖细胞(EPC)血管发生能力和在恶性胶质瘤血管新生过程中的作用.方法 应用密度梯度离心法分离新鲜人脐血的单个核细胞,接种于EGM-2培养液中培养获得EPC.取生长到第7～10天的细胞进行CD34和VEGFR-2免疫荧光双标染色.检测血管内皮生长因子(VEGF)刺激下EPC增殖活性、迁移能力和体外形成小管样结构的能力.采用人恶性胶质瘤细胞系U87在免疫缺陷小鼠进行皮下移植,于接种肿瘤后第7天经尾静脉注射EPC(每只5×103),并于接种肿瘤后第28天取材检测肿瘤微血管和EPC组织分布及定位,采用抗人CD31和抗鼠CD31免疫荧光双标记肿瘤微血管,计算人源性EPC来源的血管占肿瘤血管网的比例.结果 培养的细胞在第7～10天时可见条索样结构,生长并逐渐融合形成铺路石样排列的单层细胞,表达内皮细胞标记物CD34和VEGFR-2.在VEGF刺激下EPC具有较强的增殖活性、迁移能力和体外形成小管样结构的能力.外源性EPC能特异性归巢到异种移植瘤组织并形成新生血管,占肿瘤血管网的(18.68±1.32)%.结论 EPC在体内外具有形成血管能力,并参与异种移植瘤血管新生,提示其在恶性肿瘤血管新生过程中具有重要作用,并可能参与肿瘤微血管构筑表型异质性.
目的 觀察人臍血源性內皮祖細胞(EPC)血管髮生能力和在噁性膠質瘤血管新生過程中的作用.方法 應用密度梯度離心法分離新鮮人臍血的單箇覈細胞,接種于EGM-2培養液中培養穫得EPC.取生長到第7～10天的細胞進行CD34和VEGFR-2免疫熒光雙標染色.檢測血管內皮生長因子(VEGF)刺激下EPC增殖活性、遷移能力和體外形成小管樣結構的能力.採用人噁性膠質瘤細胞繫U87在免疫缺陷小鼠進行皮下移植,于接種腫瘤後第7天經尾靜脈註射EPC(每隻5×103),併于接種腫瘤後第28天取材檢測腫瘤微血管和EPC組織分佈及定位,採用抗人CD31和抗鼠CD31免疫熒光雙標記腫瘤微血管,計算人源性EPC來源的血管佔腫瘤血管網的比例.結果 培養的細胞在第7～10天時可見條索樣結構,生長併逐漸融閤形成鋪路石樣排列的單層細胞,錶達內皮細胞標記物CD34和VEGFR-2.在VEGF刺激下EPC具有較彊的增殖活性、遷移能力和體外形成小管樣結構的能力.外源性EPC能特異性歸巢到異種移植瘤組織併形成新生血管,佔腫瘤血管網的(18.68±1.32)%.結論 EPC在體內外具有形成血管能力,併參與異種移植瘤血管新生,提示其在噁性腫瘤血管新生過程中具有重要作用,併可能參與腫瘤微血管構築錶型異質性.
목적 관찰인제혈원성내피조세포(EPC)혈관발생능력화재악성효질류혈관신생과정중적작용.방법 응용밀도제도리심법분리신선인제혈적단개핵세포,접충우EGM-2배양액중배양획득EPC.취생장도제7～10천적세포진행CD34화VEGFR-2면역형광쌍표염색.검측혈관내피생장인자(VEGF)자격하EPC증식활성、천이능력화체외형성소관양결구적능력.채용인악성효질류세포계U87재면역결함소서진행피하이식,우접충종류후제7천경미정맥주사EPC(매지5×103),병우접충종류후제28천취재검측종류미혈관화EPC조직분포급정위,채용항인CD31화항서CD31면역형광쌍표기종류미혈관,계산인원성EPC래원적혈관점종류혈관망적비례.결과 배양적세포재제7～10천시가견조색양결구,생장병축점융합형성포로석양배렬적단층세포,표체내피세포표기물CD34화VEGFR-2.재VEGF자격하EPC구유교강적증식활성、천이능력화체외형성소관양결구적능력.외원성EPC능특이성귀소도이충이식류조직병형성신생혈관,점종류혈관망적(18.68±1.32)%.결론 EPC재체내외구유형성혈관능력,병삼여이충이식류혈관신생,제시기재악성종류혈관신생과정중구유중요작용,병가능삼여종류미혈관구축표형이질성.
Objective To investigate vasculogenic potential of endothelial progenitor cells(EPCs)derived from human umbilical cord blood and their contribution to the neovascularization of malignant glioma in vivo.Methods EPCs were isolated from human umbilical cord blood by density gradient centrifugation.After 7-10 days of culture.EPCs were investigated for CD34 and VEGFR-2 expression by direct immunofluoresent staining.The proliferative activity,migratory capability and forming capillary-like tubules were also monitored after stimulation witll VEGF(50 mg/L)in vitro.Moreover.EPCs were administered into tumor-bearing mice,and the tumor and mouse organs were examined under confocal laser scanning microscope to visualize the distribution and localization of transplanted EPCs.In order to quantity the incorporation of EPCs into tumor vessels.cryosections of the tumor tissue were double-labelled with anti-human CD31 and anti-mouse CD31.Results After 7 to 10 days of culture.EPCs assumed cobblestone-like monolayer growth pattern with nearly complete confluence,and expressed CD34 and VEGFR-2.Significant proliferative activity,increased migratory capability and forming capillary-like tubules were observed when stimulated witll VEGF.The transplanted EPCs in vivo specifically homed to solid tumor tissue and incorporated into the tumor's endothelium.Quantitative analysis revealed mat human EPCs contributed significantly to tumor neovascularization by incorporation into tumor vasculature ( 18.68±1.32)%of the total vessels.Conclusion EPCs possess the potential to form neovascular network in tumor and play a role in the phenotypical heterogeneity of tumor microvascular architecture.
