人羊膜间充质干细胞移植对荷瘤鼠C6胶质瘤生长的抑制  

Inhibitory effects of human amniotic mesenchyme stem cells on C_6 glioma cells in tumor-bearing mice

作　　者：李远[1] 焦红亮[1] 关方霞[2] 宋来君[1] 杨波[1] 胡祥 杜英[4] 李惠翔[5] 庞霞[5] 刘福生[6] 薛敬礼[7] 

机构地区：[1]郑州大学第一附属医院神经外科,河南省郑州市450052 [2]郑州大学生物工程系,河南省郑州市450001 [3]深圳市北科细胞工程研究所,广东省深圳市518000 [4]郑州大学基础医学院微生物免疫教研室,河南省郑州市450052 [5]郑州大学第一附属医院病理科,河南省郑州450052 [6]首都医科大学北京天坛医院、北京市神经外科研究所,北京市100050 [7]河南省实验动物中心,河南省郑州市450052

出　　处：《中国组织工程研究与临床康复》2008年第51期10099-10103,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(重点项目)(200702003);教育部新世纪优秀人才支持计划(2006NCET-01)~~

摘　　要：背景:实验室前期课题成果显示,脐血或羊膜间充质干细胞具有亲肿瘤及抑瘤特性。目的:探讨人羊膜间充质干细胞对C6胶质瘤细胞生长的抑制作用。设计、时间及地点:细胞学体内对照观察,于2008-06/09在郑州大学微生物与免疫教研室完成。材料:胎盘来源于郑州大学一附院妇产科健康剖宫产产妇,C6胶质瘤细胞株由北京神经外科研究所惠赠,10只BALR/c裸鼠由上海斯莱克实验动物有限责任公司提供。方法:无菌条件下取胎盘,分离部分羊膜,体外分离培养人羊膜间充质干细胞,传至第3代用于实验。10只裸鼠背部双侧皮下注射含3×106个C6胶质瘤细胞的DMEM/F12培养基,建立双侧荷瘤鼠模型。成功造模后,模型对照组于裸鼠左侧皮下注射DMEM/F12培养基50μL,细胞移植组于同一裸鼠右侧皮下注射含2×106个羊膜间充质干细胞的等量DMEM/F12培养基。主要观察指标:肿瘤生长情况,周围组织浸润及新生血管形成等病理学观察,免疫组化法检测肿瘤中细胞周期素D1的表达。结果:细胞移植后14,21d,细胞移植组肿块体积明显小于模型对照组(F=54.127,P<0.05)。模型对照组瘤体呈结节状,部分肿瘤组织中心有坏死现象,内部有新生血管形成,肌层及皮肤浸润较明显,已达腹膜;细胞移植组瘤体分叶较少,无新生血管形成,仅有局部的皮肤浸润,未见肌层浸润。模型对照组细胞周期素D1蛋白呈阳性表达,而细胞移植组表达阳性率较低。结论:人羊膜间充质干细胞可明显抑制荷瘤鼠C6胶质瘤细胞的生长,同时可抑制细胞周期素D1蛋白的异常表达。BACKGROUND： Umbilical cord blood or amniotic mesenchymal stem cells are characterized by affinity tumor and inhibiting tumor. OBJECTIVE： To explore inhibitory effects of the human amniotic mesenchyme stem cells on C6 glioma cells. DESIGN, TIME AND SETTING： The cytological in vivo controlled experiment was performed at the Department of Microblology and Immunology, Zhengzhou University from June to September 2008. MATERIALS： The amnion membrane was harvested from healthy puerperants under strict sterile conditions at the First Affiliated Hospital, Zhengzhou University. C6 glioma cells were presented by Beijing Neurosurgery Institute. Ten BALR/c nude mice were bought from Shanghai Silaike Experimental Animal Co., Ltd. METHODS： Placenta was sterily obtained to separate some amniotic membrane. Human amniotic mesenchymal stem cells were harvested in vitro. At the third passage, cells were used for this study. DMEM/F-12 culture-medium with 3×10^6 C6 glioma cells was infused into two sides of subcutaneous back of ten nude mice to induce models of mice with tumor. Following model establishment, 50 u L of DMEM/F-12 was inoculated into the mouse left side of subcutaneous tissues in the model control group. An equal volume of DMEM/F-12 containing 2×10^6 amniotic mesenchymal stem cells was injected into the right side of subcutaneous tissues in the cell transplantation group. MAIN OUTCOME MEASURES： Growth of the glioma, infiltration of surrounding tissues and neovascularization were measured. Cyclin D 1 expression in tumor was detected using immunohistochemistry. RESULTS： The tumor volume was significantly smaller in the cell transplantation group than in the model control group at days 14 and 21 following cell transplantation （F=54.127, P 〈 0.05）. In the model control group, the tumor was nodular growth, necrotic in the center, with the presence of neovascularization and obvious infiltration in the muscle and skin, even in the peritoneum. In the cell transplantation group, few sublobe slightly skin inf

关 键 词：羊膜间充质干细胞 细胞周期素D1 C6胶质瘤 

分 类 号：R394.2[医药卫生—医学遗传学]