CAJ | 학술논문

目的建立1种分离人骨肉瘤细株HMG63肿瘤干细胞新模型.方法采用无血清干细胞培养基对人骨肉瘤HMG63瘤细浮培养形成细胞球,并予阿霉素耐药处理,把耐药细胞株注入裸鼠体内致瘤,致瘤成功后,在瘤体内注射化疗药顺铂,这样使得骨肉瘤肿瘤干细胞在增殖分化的同时反复富集后,提取细胞培养筛选,这样可获得骨肉瘤干细胞和其亚型.采用逆转录-聚合酶链反应(RT-PCR)、免疫组织化学、流式细胞仪等鉴定.结果在无血清肿瘤干细胞培养基中,人HMG63细胞能有效形成悬浮生长的克隆球,流式检测结果表明克隆球低表达CD24(14％),高表达CD133 (99％),同时高表达干细胞标志物Oct-4、Nanog等.结论通过无血清悬浮球培养结合抗肿瘤耐药筛选,可以从HMG63细胞中分离出具有干细胞特征的HMG63细胞克隆.
목적 건립1충분리인골육류세주HMG63종류간세포신모형.방법 채용무혈청간세포배양기대인골육류HMG63류세부배양형성세포구,병여아매소내약처리,파내약세포주주입라서체내치류,치류성공후,재류체내주사화료약순박,저양사득골육류종류간세포재증식분화적동시반복부집후,제취세포배양사선,저양가획득골육류간세포화기아형.채용역전록-취합매련반응(RT-PCR)、면역조직화학、류식세포의등감정.결과 재무혈청종류간세포배양기중,인HMG63세포능유효형성현부생장적극륭구,류식검측결과표명극륭구저표체CD24(14％),고표체CD133 (99％),동시고표체간세포표지물Oct-4、Nanog등.결론 통과무혈청현부구배양결합항종류내약사선,가이종HMG63세포중분리출구유간세포특정적HMG63세포극륭.
Objective To isolate,purify and identify HMG63-derived cancer stem cells by suspension sphere culture combined with chemotherapy drugs.Methods HMG-63 cells were cultured in polyhema-treated dish and cancer stem cell specific medium.In this culture system,HMG63 could form spheres which were then collected and injected into the immanodeficiency mice.Cisplatin was administered every three days for five times.The enriched colon cancer xenograft tumors were cultured in cancer stem cell specific medium again to form tumor-sphere.The expression of cancer stem cells-related markers was detected by using flow cytometry.Results We obtained a CD133 +/CD24 low cell population with high expression levels of cancer stem cells-related markers,such as Oct-4,Nanog.Conclusion Sphere-formed and chemoresistant colon xenograft tumors in immunocompromised mice can enrich cancer stem cells population.Using this method we isolated HMG63-derived cancer stem cells.