胶质瘤血瘤屏障体外模型的形态学观察

MORPHOLOGICAL OBSERVATION IN VITRO MODEL OF BLOOD-TUMOR BARRIER OF GLIOMA

目的 探讨胶质瘤血瘤屏障体外模型的形态学特征。 方法 利用电镜技术观察血管内皮细胞ECV30 4 和C6 胶质瘤细胞共培养 (混合共培养、Transwell共培养、Transwell膜两面共培养 )后的内皮细胞的孔窗、内皮细胞之间的连接及内皮细胞与肿瘤细胞的相互关系、瘤细胞的“血管周足”等的形态学特征 ;并与 4例人脑胶质瘤组织标本的血瘤屏障进行比较。 结果 电镜下发现ECV30 4 细胞经与C6 细胞按 3种不同方式共培养汇合后均为无孔窗型内皮细胞 ,细胞间出现紧密连接 ;Transwell共培养的膜上C6 细胞未见伸出伪足突向膜的微孔 ;经膜两面共培养的瘤细胞则可通过Transwell的微孔突向内皮细胞侧 ,但瘤细胞未伸出伪足包绕内皮细胞或突入内皮细胞间隙 ,瘤细胞的“血管周足”不完整 ,后两者与脑胶质瘤组织标本的血瘤屏障的形态特征相类似。 结论 在体外经Transwell膜两面共培养的ECV30 4 和C6 胶质瘤细胞的系统可在一定程度模拟体内的血瘤屏障的形态学特征。

Objective To study the morphological characteristics of blood-tumor barrier(BTB)model of glioma in vitro. Methods After C 6 glioma cells/endothelia ECV 304 co-cultured mixed or in Transwell or on both sides of membrane of Transwell,the morphological characters of fenestra of endothelial cells,the junction between ECV 304 cells,the interface between C 6 cells and ECV 304 cells,and perivascular-end-feet of C 6 cells were observed under transmission electron microscope(TEM)and scanning electron microscope(SEM),and compared to BTB in 4 cases with human brain glioma. Results ECV 304 cells grown to confluence were not fenestrated,but with fomation of tight junction between ECV 304 cells after co-cultured with C 6 cells mixedly,in Transwell or on both sides of membrane of Transwell;It was not found that pseudopodia from C 6 cells as co-cultured in Transwell reaching into pore of Transwell;C 6 cells co-cultured on both sides of membrane of Transwell often sticked out to the ECV 304 cells,but with no pseudopodia from C 6 cells surrounded ECV 304 cells of penetrated the endothelia clefts.Perivascular-end-feet of C 6 cells were not integrant.These characters were similar to BTB in human brain glioma.Conclusion C 6 glioma cells/endothelia cell ECV 304 co-cultures on both sides of membrane of Transwell may simulate the morphological characters of BTB in vivo in some degree.;