Abstract: Objective  To establish the 9L/F344 rat intracerebral glioma model of brain tumor stem cells.  Methods  Rat 9L gliosarcoma stem-like cells were cultured in serum-free suspension. The expression of CD133 and nestin were tested by immunohistochemistry. A total of 48 inbredline male F344 rats were randomly divided into 2 groups, and 9L tumor sphere cells and 9L monolayer cells were respectively implanted into the right caudate nucleus of F344 rats in 2 groups. Survival time was observed and determined using the method of Kaplan-Meier survival analysis. Fourteen days after implantation or when the rats were dying, their brains were perfused and sectioned for HE staining, and CD133 and nestin were detected by immunohistochemistry.  Results  Rat 9L tumor spheres were formed with suspension culture in serum-free medium. The gliomas formed in both groups were invasive without obvious capsule. More new vessels, bleeding and necrosis could be detected in 9L tumor spheres group. The tumor cells in both groups were positive for CD133 and nestin. There was no significant difference in the expression of CD133 and nestin between 2 groups (P > 0.05, for all). According to the expression of nestin, the tumors formed by 9L tumor sphere cells were more invasive. The median survival time of the rats bearing 9L tumor sphere cells was 15 d (95%CI: 15.219-15.781), and the median survival time of the rats bearing 9L monolayer cells was 21 d (95%CI: 20.395-21.605). There was significant difference between 2 groups (χ² = 12.800, P = 0.000).  Conclusions  9L/F344 rat intracerebral glioma model of brain tumor stem cells is successfully established, which provides a glioma model for the future research.

Key words: Glioma, Neoplastic stem cells, Immunohistochemistry, Cells, cultured, Disease models, animal

摘要： 目的 建立稳定的9L/F344 大鼠脑肿瘤干细胞颅内肿瘤模型。方法 采用无血清悬浮培养法培养大鼠9L 胶质瘤细胞，立体定向法于近交系F344 大鼠右侧尾状核分别接种9L 胶质瘤细胞和9L肿瘤球细胞，观察大鼠存活状态，免疫组织化学染色检测肿瘤细胞CD133和巢蛋白表达变化。结果 9L胶质瘤细胞在无血清培养基中呈悬浮生长，形成的悬浮肿瘤细胞球表达CD133 和巢蛋白。9L 胶质瘤细胞组和9L 肿瘤细胞球组大鼠肿瘤均呈浸润性生长、无明显包膜；后者肿瘤内新生血管更丰富，出血性坏死更明显。两组肿瘤细胞均表达CD133 和巢蛋白，且组间差异无统计学意义（均P > 0.05），但9L 肿瘤细胞球组细胞更具侵袭性。接种9L 肿瘤球细胞的大鼠中位存活期为15 d（95%CI：15.219 ~ 15.781），明显短于接种9L 胶质瘤细胞大鼠的21 d（95%CI：20.395 ~ 21.605；χ² = 12.800，P = 0.000）。结论 9L/F344 大鼠脑肿瘤干细胞颅内肿瘤模型的成功建立，为进一步研究脑肿瘤干细胞提供了实验基础。

关键词: 神经胶质瘤, 肿瘤干细胞, 免疫组织化学, 细胞, 培养的, 疾病模型, 动物