人U87-MG脑胶质瘤细胞裸鼠原位移植模型的建立
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摘要
目的应用不同数量U87-MG细胞接种裸鼠脑内,建立裸鼠脑胶质瘤模型,观察其生长特性。方法采用立体定向技术,分别将3.0×10~(10)·L~(-1)、4.0×10~(10)·L~(-1)、5.0×10~(10)·L~(-1)浓度的人脑胶质瘤细胞U87-MG单细胞悬液,接种于18只♂裸鼠的右侧尾状核区,每只接种体积为5μL,另取6只裸鼠作为对照,接种同体积Hanks液。观察不同接种量实验鼠的生存状态、成瘤情况及脏器转移灶、带瘤生存期,测量肿瘤的最大径,计算肿瘤体积,将获取的全脑标本制作病理切片,行HE染色和免疫组化检查。结果各接种量的实验组成瘤率均为100%,未见颅外转移病灶,3组裸鼠的平均生存时间分别为(46.50±3.27)d、(38.50±3.28)d、(30.67±3.51)d;病理学检查符合人脑胶质瘤细胞的形态学特征和免疫表型;免疫组化GFAP和S-100蛋白呈阳性表达。结论立体定向脑内定量注射U87-MG细胞制备的人脑胶质瘤裸鼠原位移植模型成瘤率高、颅内生长稳定、颅外远隔部位转移率低,与人脑胶质瘤病理学及形态学特征相似,能为研究胶质瘤的发生、发病机制、生物学特性以及探寻有效的治疗措施提供可靠的动物模型。
Aim To establish human U87-MG glioma model in nude mice brain and to observe the characteristics of the tumor growth. Methods Human U87-MG glioma cells were cultured in vitro. 5 μL of cell suspension containing 3. 0 × 10~(10)·L~(-1),4. 0 × 10~(10)·L~(-1) and 5. 0 × 10~(10)·L~(-1) respectively was inoculated into the right caudate nucleus of 18 male nude mice brain under the guidance of stereotaxic apparatus,separately,whereas another 6 nude mice as the control group,were inoculated into the same volume of Hanks solution. The moving and survival state of rats with gliomas were observed. The examinations of the tumors formation,volumes,metastasis and histopathology were performed and the obtained brain samples were stained with HE and immunohistochemistry. Results All the tested rats of different inoculation doses developed brain tumors without extracranial metastasis. The mean survival time of three groups was( 46. 50 ± 3. 27) d,( 38. 50 ± 3. 28) d and( 30. 67 ± 3. 51) d,respectively. The tumors showed the similar morphological features and immunophenotype to human glioma. There was positive expression of GFAP and S-100 in the tumors. Conclusions The orthotopic implantation model of human U87-MG glioma,by inoculating quantitative U87-MG cells stereotaxically into the brains of the nude mice,is successfully established with 100 yield of intracranial tumor and no extracranial growth extension.It resembles the histopathological and morphological features of human glioma,which can be used as a reliable animal model for the study of the tumorigenesis,pathogenesis,biological characteristics and therapy of glioma.
Aim To establish human U87-MG glioma model in nude mice brain and to observe the characteristics of the tumor growth. Methods Human U87-MG glioma cells were cultured in vitro. 5 μL of cell suspension containing 3. 0 × 10~(10)·L~(-1),4. 0 × 10~(10)·L~(-1) and 5. 0 × 10~(10)·L~(-1) respectively was inoculated into the right caudate nucleus of 18 male nude mice brain under the guidance of stereotaxic apparatus,separately,whereas another 6 nude mice as the control group,were inoculated into the same volume of Hanks solution. The moving and survival state of rats with gliomas were observed. The examinations of the tumors formation,volumes,metastasis and histopathology were performed and the obtained brain samples were stained with HE and immunohistochemistry. Results All the tested rats of different inoculation doses developed brain tumors without extracranial metastasis. The mean survival time of three groups was( 46. 50 ± 3. 27) d,( 38. 50 ± 3. 28) d and( 30. 67 ± 3. 51) d,respectively. The tumors showed the similar morphological features and immunophenotype to human glioma. There was positive expression of GFAP and S-100 in the tumors. Conclusions The orthotopic implantation model of human U87-MG glioma,by inoculating quantitative U87-MG cells stereotaxically into the brains of the nude mice,is successfully established with 100 yield of intracranial tumor and no extracranial growth extension.It resembles the histopathological and morphological features of human glioma,which can be used as a reliable animal model for the study of the tumorigenesis,pathogenesis,biological characteristics and therapy of glioma.
引文
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[2]黄强.胶质瘤干细胞研究的现状与未来[J].中华神经医学杂志,2007,6(9):865-8.[2]Huang Q.Glioma stem cells:current situation and prospects[J].Chin J Neuromed,2007,6(9):865-8.
[3]Antunes L,Angioi-Duprez K S,Bracard S R,et al.Analysis of tissue chimerism in nude mouse brain and abdominal xenograft models of human glioblastoma multiforme:what does it tell us about the models and about glioblastoma biology and therapy[J]?J Histochem Cytochem,2000,48(6):847-58.
[4]Brekken C,Bruland S,de Lange Davies C.Interstitial fluid pressure in human osteosarcoma xenografts:significance of implantation site and the response to intratumoral injection of hyaluronidase[J].Anticancer Res,2000,20(5B):3503-12.
[5]崔白苹,陈平,孙安阳.脑转移瘤动物模型的制备方法及研究进展[J].中国药理学通报,2016,32(3):304-9.[5]Cui B P,Chen P,Sun A Y.Animal models of brain metastasis:preparation methods and research progress[J].Chin Pharmacol Bull,2016,32(3):304-9.
[6]徐承平,卞修武.人CHG-5胶质瘤细胞裸鼠原位移植模型的建立及其生物学特性分析[J].第三军医大学学报,2003,25(4):287-90.[6]Xu C P,Bian X W.Establishment of orthotopic implantation model of human CHG-5 glioma cell line in nude mice and analysis of its biological features[J].2003,25(4):287-90.
[7]余永强,陈骏,钱银锋,等.立体定向建立人鼠C6脑胶质瘤模型[J].中国药理学通报,2003,19(2):222-5.[7]Yu Y Q,Chen J,Qian Y F,et al.Establishment of rat C6 glioma model by intracranial injection[J].Chin Pharmacol Bull,2003,19(2):222-5.