Quantitative analysis of glioma cell invasion by confocal laser scanning microscopy in a novel brain slice model
Quantitative analysis of glioma cell invasion by confocal laser scanning microscopy in a novel brain slice model
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DOI：
10.1006/bbrc.2000.2332
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发表时间：
2000-03-16
影响因子：
3.1
通讯作者：
Yoshimine, T
中科院分区：
文献类型：
--
作者：
Matsumura, H;Ohnishi, T;Yoshimine, T
为了定量分析胶质瘤细胞在器官型脑切片培养中迁移的空间范围，我们利用激光共聚焦扫描显微镜（CLSM）建立了一种新的侵袭模型。CLSM：不仅可以对活体脑切片中人类T98G胶质瘤细胞的侵袭模式进行三维可视化，而且可以在几周内对侵袭过程进行连续分析。T98G：胶质瘤球体与脑切片共培养24小时后，胶质瘤细胞脱离球体，自发地在脑切片表面继续迁移，同时向更深的部位弥漫性侵入。免疫组化分析显示，这些迁移的胶质瘤细胞对基质金属蛋白酶(MMP)-2和-9的免疫染色比留在植入部位的肿瘤球体强得多。用MMPs特异性抑制剂1,10-菲罗啉处理T98G胶质瘤球体，不仅能显著抑制细胞在脑切片表面的迁移，还能显著抑制胶质瘤细胞向脑切片的侵袭。目前使用CLSM建立的胶质瘤侵袭模型，可以在空间和序列上分析胶质瘤细胞在几周内的活脑切片的侵袭程度，使其成为研究胶质瘤在最类似于正常大脑的条件下侵袭的细胞和分子机制的非常有用的工具。(C) 2000年学术出版社。
To quantitatively analyze the spatial extent of glioma cell migration in an organotypic brain slice culture, we developed a new invasion model with the aid of confocal laser scanning microscopy (CLSM). CLSM: allowed not only for three-dimensional visualization of the invasive pattern of human T98G glioma cells in the living brain slice but also for serial analysis of the invasive process over several weeks. Twenty-four hours after the T98G: glioma spheroid was initiated to coculture with a brain slice, the glioma cells detached themselves from the spheroid and spontaneously continued to migrate on the surface of the brain slice, while they diffusely invaded into the slice by migrating to a deeper site. Immunohistochemical analysis revealed that these migrating glioma cells much more strongly immunostained for matrix metalloproteinase (MMP)-2 and -9 than the tumor spheroid which remained at the implanted site. Treatment of the T98G glioma spheroid with 1,10-phenanthroline, a specific inhibitor of MMPs, significantly inhibited not only the cell migration on the surface of the brain slice but also the invasion of the glioma cells into the slice. The present version of the glioma invasion model using CLSM makes it possible to spatially and serially analyze the extent of glioma cell invasion in the living brain slice for several weeks, making it a very useful tool for investigating the cellular and molecular mechanisms of glioma invasion under conditions most analogous to those of normal brains in vivo. (C) 2000 Academic Press.