中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2012年
12期
1200-1203
,共4页
刘天助%颜晓慧%汪鑫%白义凤%廖红展%苑召虎%吴炳义%邱胜聪
劉天助%顏曉慧%汪鑫%白義鳳%廖紅展%苑召虎%吳炳義%邱勝聰
류천조%안효혜%왕흠%백의봉%료홍전%원소호%오병의%구성총
肿瘤干细胞%球囊%蛋清包埋%二次包埋
腫瘤榦細胞%毬囊%蛋清包埋%二次包埋
종류간세포%구낭%단청포매%이차포매
Cancer stem cell%Tumor sphere%Egg-white-paraffin embedding technique%Secondary microencapsulation
目的 探索一种新的能长期、高效保存并研究肿瘤干细胞球囊的技术.方法 从恶性胶质瘤原代肿瘤细胞中培养出肿瘤干细胞.将第5代肿瘤干细胞球囊经蛋清石蜡两次包埋、切片后,行HE染色、免疫组织化学染色和免疫组织荧光染色,并与传统的肿瘤干细胞球囊包埋技术及球囊免疫组织荧光染色实验进行比较.结果 应用蛋清石蜡两次包埋处理的球囊切片经HE染色后,球囊分布均匀、结构完整,胞核、胞浆颜色鲜艳、对比度高.球囊切片的免疫组织化学染色和免疫组织荧光染色能准确定位球囊中阳性细胞和阳性蛋白表达位点,可进行阳性细胞及蛋白的半定量分析.结论 肿瘤干细胞球囊蛋清石蜡两次包埋切片技术能提高实验效率,减少实验误差,节省成本,优于传统球囊包埋技术及传统球囊免疫荧光技术.
目的 探索一種新的能長期、高效保存併研究腫瘤榦細胞毬囊的技術.方法 從噁性膠質瘤原代腫瘤細胞中培養齣腫瘤榦細胞.將第5代腫瘤榦細胞毬囊經蛋清石蠟兩次包埋、切片後,行HE染色、免疫組織化學染色和免疫組織熒光染色,併與傳統的腫瘤榦細胞毬囊包埋技術及毬囊免疫組織熒光染色實驗進行比較.結果 應用蛋清石蠟兩次包埋處理的毬囊切片經HE染色後,毬囊分佈均勻、結構完整,胞覈、胞漿顏色鮮豔、對比度高.毬囊切片的免疫組織化學染色和免疫組織熒光染色能準確定位毬囊中暘性細胞和暘性蛋白錶達位點,可進行暘性細胞及蛋白的半定量分析.結論 腫瘤榦細胞毬囊蛋清石蠟兩次包埋切片技術能提高實驗效率,減少實驗誤差,節省成本,優于傳統毬囊包埋技術及傳統毬囊免疫熒光技術.
목적 탐색일충신적능장기、고효보존병연구종류간세포구낭적기술.방법 종악성효질류원대종류세포중배양출종류간세포.장제5대종류간세포구낭경단청석사량차포매、절편후,행HE염색、면역조직화학염색화면역조직형광염색,병여전통적종류간세포구낭포매기술급구낭면역조직형광염색실험진행비교.결과 응용단청석사량차포매처리적구낭절편경HE염색후,구낭분포균균、결구완정,포핵、포장안색선염、대비도고.구낭절편적면역조직화학염색화면역조직형광염색능준학정위구낭중양성세포화양성단백표체위점,가진행양성세포급단백적반정량분석.결론 종류간세포구낭단청석사량차포매절편기술능제고실험효솔,감소실험오차,절성성본,우우전통구낭포매기술급전통구낭면역형광기술.
Objective To explore a new long-term efficient embedding technique of tumor spheres.Methods Tumor spheres,mainly composed of cancer stem cells,were cultured from glioblastoma tissues.The fifth generation of tumor spheres was chosen for egg-white-paraffin embedding and section.Then,those tumor sphere slices were observed by HE staining,immunohistochemistry and immunofluorescence staining.And the immunofluorescence results of these tumor sphere slices were compared with those tumor spheres kept with traditional methods.Results Immunofluorescence results showed that tumor spheres kept with traditional methods looked blurry,and the positive cells and the positive protein expression sites in the cells could not be displayed.HE staining demonstrated that the tumor sphere slices had well-distributed intact spheres and coloring cells with high karyoplasm contrast.Immunohistochemistry and immunofluorescence staining of tumor sphere slices showed clear background,from which positive cells and positive locus could be easily displayed; therefore,semi-quantitative analysis of the positive cells could be performed.Conclusion Egg-white-paraffin embedding technique of the tumor sphere slices can reduce experimental errors and cut down the costs,which enjoys its advantage as compared with traditional embedding technique of the tumor sphere slices.