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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: The results of pre-laboratory research show that cord blood or amniotic mesenchymal stem cells have the characteristics of pro-tumor and anti-tumor. Objective: To investigate the inhibitory effect of human amniotic mesenchymal stem cells on the growth of C6 glioma cells. DESIGN, TIME AND SETTING: The cytology in vivo control study was performed at the Department of Microbiology and Immunology, Zhengzhou University from June to September 2008. MATERIALS: The placenta was derived from a healthy cesarean section in obstetrics and gynecology of First Affiliated Hospital of Zhengzhou University. The C6 glioma cell line was kindly donated by Beijing Institute of Neurosurgery. Ten BALR / c nude mice were provided by Shanghai Slack Laboratory Animal Co., Ltd. . Methods: The placenta was taken under aseptic conditions and part of amniotic membrane was isolated. Human amniotic mesenchymal stem cells were isolated and cultured in vitro. The cells were passaged to passage 3 for experiment. Ten nude mice were injected subcutaneously with DMEM / F12 medium containing 3 × 106 C6 glioma cells on both sides of the back to establish bilateral tumor bearing mice model. After successful modeling, the model control group was subcutaneously injected with 50 μL of DMEM / F12 medium on the left side of the nude mice and the same group of DMEM / F12 culture medium containing 2 × 106 amniotic mesenchymal stem cells base. MAIN OUTCOME MEASURES: Tumor growth, peripheral tissue infiltration and neovascularization pathological observation, immunohistochemical detection of tumor cell cyclin D1 expression. RESULTS: At 14 and 21 days after cell transplantation, the volume of tumor in the cell transplantation group was significantly smaller than that in the model control group (F = 54.127, P <0.05). The control group had nodules in the tumor, necrosis in the center of some tumor tissues, neovascularization in the internal part, obvious infiltration of the muscle layer and skin, and reached the peritoneum. There were fewer tumor lobes and no neovascularization in the cell transplantation group , Only partial skin infiltration, no muscle invasion. The expression of cyclin D1 protein in the model control group was positive, but the positive rate in the cell transplantation group was lower. Conclusion: Human amniotic mesenchymal stem cells can significantly inhibit the growth of C6 glioma cells and inhibit the abnormal expression of cyclin D1 protein.