中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
14期
2641-2648
,共8页
曲绍政%李书忠%高甲科%张金锋
麯紹政%李書忠%高甲科%張金鋒
곡소정%리서충%고갑과%장금봉
干细胞%干细胞学术探讨%骨肿瘤%骨肉瘤%肿瘤干细胞%阿霉素%长春新碱%CD133%Nanog%Stro-1%悬浮细胞
榦細胞%榦細胞學術探討%骨腫瘤%骨肉瘤%腫瘤榦細胞%阿黴素%長春新堿%CD133%Nanog%Stro-1%懸浮細胞
간세포%간세포학술탐토%골종류%골육류%종류간세포%아매소%장춘신감%CD133%Nanog%Stro-1%현부세포
stem cel s%stem cel academic discussion%bone tumors%osteosarcoma%tumor stem cel s%doxorubicin%vincristine%CD133%Nanog%Stro-1%suspended cel s
背景:骨肉瘤是最常见的骨原发性恶性肿瘤,其发生可能与骨肉瘤干细胞有关.目的:评估骨肉瘤干细胞分离、培养和鉴定方法以及其相关肿瘤标记物的表达.方法:在无血清条件下以及无血清联合抗肿瘤药物的条件下应用免疫磁珠分选法对骨肉瘤干细胞进行分离、培养,分选出 Stro-1阳性、CD133阳性骨肉瘤干细胞,采用免疫荧光染色、蛋白质印迹法等检测骨肉瘤肿瘤干细胞标记物 CD133、Oct3/4以及 Nanog 等的表达水平以及致瘤性能.结果与结论:骨肉瘤干细胞在接种培养2-10 d 后形成悬浮细胞球,增殖潜伏期约为24 h,Stro-1阳性干细胞能够形成悬浮细胞球,Stro-1阴性细胞则不能形成悬浮细胞球.此外,骨肉瘤干细胞还能高表达Oct3/4、Nanog 和 CD133等,CD133阳性骨肉瘤干细胞高表达 CD133分子,侵袭力更强,而 CD133阴性细胞则不能表达 CD133分子,侵袭力相对较弱.塞来昔布对骨肉瘤干细胞的形成具有一定程度的抑制作用,能够降低肿瘤新生血管中血管内皮生长因子的表达.
揹景:骨肉瘤是最常見的骨原髮性噁性腫瘤,其髮生可能與骨肉瘤榦細胞有關.目的:評估骨肉瘤榦細胞分離、培養和鑒定方法以及其相關腫瘤標記物的錶達.方法:在無血清條件下以及無血清聯閤抗腫瘤藥物的條件下應用免疫磁珠分選法對骨肉瘤榦細胞進行分離、培養,分選齣 Stro-1暘性、CD133暘性骨肉瘤榦細胞,採用免疫熒光染色、蛋白質印跡法等檢測骨肉瘤腫瘤榦細胞標記物 CD133、Oct3/4以及 Nanog 等的錶達水平以及緻瘤性能.結果與結論:骨肉瘤榦細胞在接種培養2-10 d 後形成懸浮細胞毬,增殖潛伏期約為24 h,Stro-1暘性榦細胞能夠形成懸浮細胞毬,Stro-1陰性細胞則不能形成懸浮細胞毬.此外,骨肉瘤榦細胞還能高錶達Oct3/4、Nanog 和 CD133等,CD133暘性骨肉瘤榦細胞高錶達 CD133分子,侵襲力更彊,而 CD133陰性細胞則不能錶達 CD133分子,侵襲力相對較弱.塞來昔佈對骨肉瘤榦細胞的形成具有一定程度的抑製作用,能夠降低腫瘤新生血管中血管內皮生長因子的錶達.
배경:골육류시최상견적골원발성악성종류,기발생가능여골육류간세포유관.목적:평고골육류간세포분리、배양화감정방법이급기상관종류표기물적표체.방법:재무혈청조건하이급무혈청연합항종류약물적조건하응용면역자주분선법대골육류간세포진행분리、배양,분선출 Stro-1양성、CD133양성골육류간세포,채용면역형광염색、단백질인적법등검측골육류종류간세포표기물 CD133、Oct3/4이급 Nanog 등적표체수평이급치류성능.결과여결론:골육류간세포재접충배양2-10 d 후형성현부세포구,증식잠복기약위24 h,Stro-1양성간세포능구형성현부세포구,Stro-1음성세포칙불능형성현부세포구.차외,골육류간세포환능고표체Oct3/4、Nanog 화 CD133등,CD133양성골육류간세포고표체 CD133분자,침습력경강,이 CD133음성세포칙불능표체 CD133분자,침습력상대교약.새래석포대골육류간세포적형성구유일정정도적억제작용,능구강저종류신생혈관중혈관내피생장인자적표체.
@@@@BACKGROUND: Osteosarcoma is the most common bone primary malignant tumors, and the occurrence of osteosarcoma may relate with osteosarcoma stem cel s. OBJECTIVE: To evaluate the isolation, culture and identification method of osteosarcoma stem cel s, to investigate the expression of relative tumor markers. METHODS: The osteosarcoma stem cel s were isolated and cultured with magnetic activated cel sorting method under serum-free condition and serum-free joint antineoplastic condition, in order to sort Stro-1 positive and CD133 positive osteosarcoma stem cel s. The expression level and tumorigenic properties of CD133, Oct3 / 4 and Nanog markers of osteosarcoma cancer stem cel s were tested with immunofluorescence staining and Western blotting methods. RESULTS AND CONCLUSION: Suspended cel condensation could be seen in the osteosarcoma stem cel s after culture for 2-10 days, the proliferation incubation period was about 24 hours, and the Stro-1 positive stem cel s could form the suspended cel condensation, while the Stro-1 negative stem cel s could not form the suspended cel condensation. In addition, osteosarcoma stem cel s could highly express Oct3/4, Nanog and CD133, the CD133-positive osteosarcoma stem cel s could highly express CD133 molecule with strong invasiveness, while the CD133 negative cel s could not express CD133 molecule and the invasiveness was weak. Celecoxib could inhibit the formation of osteosarcoma stem cel s to some extent, and could reduce the expression of tumor angiogenic blood vessels and vascular endothelial growth factor.
