Nygaard S J, Haugland H K, Laerum O D, Lund-Johansen M, Bjerkvig R, Tysnes O B
Department of Pathology, The Gade Institute, University of Bergen, Haukeland Hospital, Norway.
J Neurosurg. 1998 Sep;89(3):441-7. doi: 10.3171/jns.1998.89.3.0441.
The goal of this study was to evaluate whether there is any relationship between survival of patients with brain tumor and tumor proliferation or tumor invasion in vitro.
Samples of freshly resected brain tumors from 14 patients with glioblastoma multiforme (GBM) were directly grown as three-dimensional multicellular spheroids. The tumor spheroids were cocultured with fetal rat brain cell aggregates (BCAs), used to represent an organotypical normal brain tissue model. Before the coculture, the tumor spheroids and the BCAs were stained with two different carbocyanine dyes, 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) and 3,3'-dioctadecycloxacarbocyanine perchlorate (DiO), respectively. During the coculture, confocal laser scanning microscopy allowed a sequential analysis of tumor cell invasion by visualizing dynamic aspects of the invasive process. Single cocultures were examined at three different time points (24, 48, and 96 hours). During the observation period there was a change in the structural morphology of the cocultures, with a progressive decrease in BCA volume. Furthermore, the scanning confocal micrographs revealed a bidirectional movement of tumor cells and normal cells into brain and tumor tissue, respectively. It is also shown that there is a considerable variation in the rate of BCA destruction in cocultures of glioma spheroids generated directly from biopsy specimens. This variation is seen both between spheroids generated from the same biopsy as well as between spheroids that are grown from different biopsy specimens. Cell proliferation measured by Ki-67 immunohistochemical analysis of biopsy samples obtained in the same patients revealed a correlation between tumor cell proliferation and tissue destruction of the BCAs, as determined by a reduction in BCA volume (p = 0.0338). No correlation was found when survival was related to the same parameters (p > 0.05).
The present work provides a model for quick and efficient assessment of dynamic interactions between tumor and normal brain tissue shortly after surgery.
本研究的目的是评估脑肿瘤患者的生存期与肿瘤增殖或体外肿瘤侵袭之间是否存在任何关联。
从14例多形性胶质母细胞瘤（GBM）患者新鲜切除的脑肿瘤样本直接培养成三维多细胞球体。肿瘤球体与胎鼠脑细胞聚集体（BCA）共培养，后者用于代表器官典型正常脑组织模型。在共培养前，肿瘤球体和BCA分别用两种不同的碳菁染料，即1,1'-二辛基-3,3,3',3'-四甲基吲哚碳菁高氯酸盐（DiI）和3,3'-二辛基氧杂碳菁高氯酸盐（DiO）染色。在共培养过程中，共聚焦激光扫描显微镜通过观察侵袭过程的动态方面，对肿瘤细胞侵袭进行了连续分析。在三个不同时间点（24、48和96小时）检查单个共培养物。在观察期内，共培养物的结构形态发生了变化，BCA体积逐渐减小。此外，扫描共聚焦显微照片显示肿瘤细胞和正常细胞分别向脑和肿瘤组织双向移动。还表明，直接从活检标本生成的胶质瘤球体共培养物中BCA破坏率存在相当大的差异。这种差异在来自同一活检的球体之间以及来自不同活检标本的球体之间均可见。通过对同一患者活检样本进行Ki-67免疫组化分析测量的细胞增殖显示，肿瘤细胞增殖与BCA的组织破坏之间存在相关性，BCA体积减小确定了这种相关性（p = 0.0338）。当生存期与相同参数相关时未发现相关性（p > 0.05）。
本研究提供了一个模型，用于在手术后不久快速有效地评估肿瘤与正常脑组织之间的动态相互作用。
