Valadez J Geraldo, Sarangi Anuraag, Lundberg Christopher J, Cooper Michael K
Department of Neurology, Vanderbilt University Medical Center.
J Vis Exp. 2014 Jan 14(83):e50865. doi: 10.3791/50865.
Malignant gliomas constitute a heterogeneous group of highly infiltrative glial neoplasms with distinct clinical and molecular features. Primary orthotopic xenografts recapitulate the histopathological and molecular features of malignant glioma subtypes in preclinical animal models. To model WHO grades III and IV malignant gliomas in transplantation assays, human tumor cells are xenografted into an orthotopic site, the brain, of immunocompromised mice. In contrast to secondary xenografts that utilize cultured tumor cells, human glioma cells are dissociated from resected specimens and transplanted without prior passage in tissue culture to generate primary xenografts. The procedure in this report details tumor sample preparation, intracranial transplantation into immunocompromised mice, monitoring for tumor engraftment and tumor harvesting for subsequent passage into recipient animals or analysis. Tumor cell preparation requires 2 hr and surgical procedure requires 20 min/animal.
恶性胶质瘤是一组异质性的、具有高度浸润性的神经胶质瘤,具有独特的临床和分子特征。原发性原位异种移植在临床前动物模型中重现了恶性胶质瘤亚型的组织病理学和分子特征。为了在移植试验中模拟世界卫生组织(WHO)III级和IV级恶性胶质瘤,将人类肿瘤细胞异种移植到免疫缺陷小鼠的原位部位——大脑中。与利用培养的肿瘤细胞的二次异种移植不同,人类胶质瘤细胞从切除的标本中解离出来,未经组织培养传代就进行移植,以产生原发性异种移植。本报告中的程序详细介绍了肿瘤样本制备、免疫缺陷小鼠的颅内移植、肿瘤植入监测以及肿瘤采集,以便随后移植到受体动物中或进行分析。肿瘤细胞制备需要2小时,手术过程每只动物需要20分钟。