CAJ | 학술논문

目的从胶质瘤组织中分离和培养出肿瘤干细胞,并初步探讨其生长特性. 方法收集脑胶质瘤手术标本并获取细胞,用含有表皮生长因子(EGF)、白血病细胞抑制因子(LIE)和碱性成纤维生长因子(bFGF)的无血清培养液原代培养,再经免疫磁珠分离得到CD133+细胞.并用免疫细胞化学技术检测CD133、NSE和GFAP在细胞中的表达以鉴定CD133+细胞,比较不同恶性级别胶质瘤组织的CD133+细胞生长情况,并用CCK8法比较CD133+和CD133-细胞对替尼泊苷(VM-26)的耐药性. 结果从胶质瘤组织中成功分选获得CD133+细胞,这些细胞能自我更新,增殖,并分化成NSE+和GFAP+的细胞.恶性度高的胶质瘤组织中CD133+细胞生长速度明显比低级别中的CD133+细胞快,且CD133+细胞在含有VM-26培养基中的存活细胞数显著多于CD133-细胞(P＜0.05). 结论胶质瘤组织中存在肿瘤干细胞,这类细胞具有很强的耐药性,高恶性度胶质瘤组织中的CD133+细胞具有更强的增殖能力.
목적 종효질류조직중분리화배양출종류간세포,병초보탐토기생장특성. 방법 수집뇌효질류수술표본병획취세포,용함유표피생장인자(EGF)、백혈병세포억제인자(LIE)화감성성섬유생장인자(bFGF)적무혈청배양액원대배양,재경면역자주분리득도CD133+세포.병용면역세포화학기술검측CD133、NSE화GFAP재세포중적표체이감정CD133+세포,비교불동악성급별효질류조직적CD133+세포생장정황,병용CCK8법비교CD133+화CD133-세포대체니박감(VM-26)적내약성. 결과 종효질류조직중성공분선획득CD133+세포,저사세포능자아경신,증식,병분화성NSE+화GFAP+적세포.악성도고적효질류조직중CD133+세포생장속도명현비저급별중적CD133+세포쾌,차CD133+세포재함유VM-26배양기중적존활세포수현저다우CD133-세포(P＜0.05). 결론 효질류조직중존재종류간세포,저류세포구유흔강적내약성,고악성도효질류조직중적CD133+세포구유경강적증식능력.
Objective To isolate and culture brain tumor stem cells (BTSCs) from glioma tissues and explore the biological characteristics of BTSCs. Methods Different grade glioma tissues were obtained from 20 clinical cases. After tumors were dissociated, the sample was triturated into the single cells and then filtered. The primary glioma cells were collected and cultured in the DMEM/F12 medium containing epidermal growth factor (EGF), leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF), in order to promote the proliferation of BTSCs. CD133 + cells were separated by immunomagnetic bead method and identified by testing the expressions of CD133, NSE and GFAP using immunocytochemistry. CCK8 was employed to assay the proliferating situation of CD133+ cells in the different grade gliomas, and to compare the drug resistance between the CD133+ and CD133- cells in the medium containing VM-26. Results CD133+ cells were successfully separated from glioma tissues.CD133+ cells proliferated by self-renewal, then differentiated into NSE+ cells and GFAP+ ones respectively. CD133+ cells in the high grade gliomas showed the faster generation than the ones in the low grade gliomas. CD133+ cells survived more easily than the CD133- cells in the medium containing VM-26. Conclusions BTSCs exist in the glioma tissues, and possess the more tolerant to the VM-26.CD133+ cells in the high grade glioma can proliferate much more easily.