Chicoine M R, Silbergeld D L
Department of Neurological Surgery, Washington University School of Medicine, St. Louis, Missouri, USA.
J Neurosurg. 1995 Oct;83(4):665-71. doi: 10.3171/jns.1995.83.4.0665.
To characterize rat glioma cell invasion, 2 x 10(6) fluorophore-labeled or transfection-labeled C6 rat glioma cells were implanted in the rat frontal lobe. Eighty percent of the rats implanted formed bulk tumors (3-4 mm in diameter). Two weeks after implantation, fluorescence microscopy revealed single tumor cells in sites over 16 mm from the bulk brain tumor. Tumor cells distant from the bulk tumor remained single without mass formation and invaded primarily along white matter tracts. Two weeks after tumor implantation, three cell lines were created from each brain by disaggregation and initiation in culture of 1) bulk tumor, 2) contralateral hemisphere, and 3) cerebellum; all disaggregated specimens generated viable cultures. Cells cultured from the contralateral hemisphere were morphologically indistinguishable from cells from the bulk tumor and from the original C6 cell line. Cells cultured from the cerebellum were morphologically quite distinct from the C6 cell line. Cells from disaggregated specimens obtained from the tumor, contralateral hemisphere, and cerebellum were implanted in the frontal lobe of naive rats to test tumorgenicity. Bulk tumor formed in 58% of the rats implanted with specimens from tumor, in 75% of the rats implanted with specimens from contralateral hemisphere, and in only 12.5% of the rats implanted with specimens from the cerebellar hemispheres. Experiments using C6 cells labeled by transfection with the p3' ss DNA vector prior to implantation confirmed that the cells cultured from the contralateral hemisphere were derived from the implanted C6 cells. Experiments with C6 cells anchored in agar served to verify that movement to the contralateral hemisphere was secondary to parenchymal invasion rather than dispersion in the cerebrospinal fluid.
为了表征大鼠胶质瘤细胞的侵袭情况，将2×10⁶个荧光团标记或转染标记的C6大鼠胶质瘤细胞植入大鼠额叶。80%植入的大鼠形成了大块肿瘤（直径3 - 4毫米）。植入两周后，荧光显微镜检查发现在距大块脑肿瘤超过16毫米的部位有单个肿瘤细胞。远离大块肿瘤的肿瘤细胞保持单个状态，没有形成肿块，主要沿白质束侵袭。肿瘤植入两周后，通过对每个大脑进行解离并在培养中起始培养，从以下三个部位创建了三个细胞系：1）大块肿瘤，2）对侧半球，3）小脑；所有解离的标本都产生了活细胞培养物。从对侧半球培养的细胞在形态上与来自大块肿瘤的细胞以及原始C6细胞系的细胞没有区别。从小脑培养的细胞在形态上与C6细胞系有很大不同。将从肿瘤、对侧半球和小脑获得的解离标本中的细胞植入未处理大鼠的额叶以测试致瘤性。在植入来自肿瘤标本的大鼠中，58%形成了大块肿瘤；在植入来自对侧半球标本的大鼠中，75%形成了大块肿瘤；而在植入来自小脑半球标本的大鼠中，只有12.5%形成了大块肿瘤。在植入前用p3' ss DNA载体转染标记的C6细胞进行的实验证实，从对侧半球培养的细胞源自植入的C6细胞。用锚定在琼脂中的C6细胞进行的实验用于验证向对侧半球的移动是实质侵袭的结果，而不是在脑脊液中的扩散。
