Ohnishi T, Matsumura H, Izumoto S, Hiraga S, Hayakawa T
Department of Neurosurgery, Osaka University Medical School, Suita, Japan.
Cancer Res. 1998 Jul 15;58(14):2935-40.
A novel model of glioma cell invasion was established by using an organotypic culture of rat brains. Brain slices prepared from 2-day-old rat neonates were maintained in a culture at the interface between air and the culture medium. The slices were placed on double-layered membranes consisting of a polycarbonate membrane with 8-microm pores and a membrane with 0.4-microm pores. The organotypic cytoarchitecture of the cultured brain slices remained well preserved, and the neuronal viability was kept intact for over 2 months. When C6 glioma spheroids were cocultured with the brain slices, the tumor cells migrated in a scattered fashion around the spheroids. Exogenous L1 or glioma motility factor I strongly stimulated the cell migration, whereas fibronectin, tenascin, and glioma motility factor II had little or no effect. When C6 glioma cells placed on the brain slices were incubated while being stimulated by L1-transfected fibroblast cells for 2 days, many more tumor cells invaded and reached the bottom of the upper membrane. This L1-stimulated glioma cell invasion into brain slices was significantly inhibited by an anti-L1 antibody. Our novel invasion model, which mimics the in vivo conditions of the central nervous system, may make it possible to analyze actual events of glioma cell invasion in normal brains in situ.
通过使用大鼠脑的器官型培养建立了一种新的胶质瘤细胞侵袭模型。从2日龄大鼠新生儿制备的脑片维持在空气与培养基界面的培养中。脑片放置在由具有8微米孔的聚碳酸酯膜和具有0.4微米孔的膜组成的双层膜上。培养的脑片的器官型细胞结构保持良好保存，并且神经元活力在2个月以上保持完整。当C6胶质瘤球体与脑片共培养时，肿瘤细胞以分散的方式在球体周围迁移。外源性L1或胶质瘤运动因子I强烈刺激细胞迁移，而纤连蛋白、腱生蛋白和胶质瘤运动因子II几乎没有或没有影响。当放置在脑片上的C6胶质瘤细胞在L1转染的成纤维细胞刺激下孵育2天时，更多的肿瘤细胞侵入并到达上膜的底部。这种L1刺激的胶质瘤细胞向脑片的侵袭被抗L1抗体显著抑制。我们模拟中枢神经系统体内条件的新型侵袭模型可能使原位分析正常脑中胶质瘤细胞侵袭的实际事件成为可能。
