Preparation of aniti-tumor tissue endothelium antibodies and its application of cancer-missle therapy

fitc标记的大分子葡聚糖（Mw=70,000）通过用小鼠黑色素瘤B16制备的条件培养基培养5天的牛主动脉内皮细胞（BAEC）单层膜的通透性增加。而在正常培养基中培养至融合的BAEC，用B16条件培养基（B16- cm）处理30min后，通透性没有增加。B16-CM还能增加牛静脉和人脐静脉内皮层的通透性，但没有增加上皮层的通透性。B16-CM未改变BAEC上F-actin的分布和含量。在B16-CM存在下培养5 d的BAEC脱离培养皿，然后按细胞密度的五分之一接种于室中。在正常培养基中培养5天后，BAEC生长至融合，其渗透性增加。这些结果表明B16-CM在不降低F-actin的情况下，不可逆地增加了内皮细胞的通透性，肿瘤细胞分泌的可溶性因子参与了体内肿瘤血管高通透性结构的构建。采用Percoll梯度离心附速分离技术分离大鼠KMT-17纤维肉瘤源性内皮细胞，观察其培养性能。原代培养的肿瘤源性内皮细胞（TEC）显示血管紧张素转换酶活性，因子viii相关抗原染色阳性，并在Matrigel上形成典型的毛细血管样结构。原代培养的TEC单层比正常组织源性内皮细胞（主动脉、腔静脉和附睾脂肪毛细血管）单层在fitc -葡聚糖扩散（分子量70000）上表现出更大的渗透性。与脂肪来源的毛细血管内皮细胞相比，白细胞对TEC的粘附减少。这些特征与肿瘤血管内皮相似，并且在原代和第一代细胞培养中都观察到，但在第四代细胞培养中没有观察到。我们的研究结果表明，原代或传代培养的TEC可用于研究肿瘤内皮细胞的生理特性。少

The permeation of macromolecular FITC-labeled dextran (Mw=70,000) through bovine aortic endothelial cells (BAEC) monolayr, which were cultured for 5 days with conditioned medium prepared from mouse melanoma B16, was increased. However, when BAEC,which were cultured with normal medium until confluent, were treated with B16 conditioned medium (B16-CM) for 30 min, the permeability did not increase. The B16-CM also increased the permeability of the endothelial monolayrs of bovine veins and the human umbilical vein, but did not increase that of the epithelial monolayr. The B16-CM did not alter the distribution or content of F-actin on the BAEC.BAEC cultured in the presence of B16-CM for 5 days were detached from the dish, and then seeded into a chamber at one-fifth of confluent cell density. After 5 days of culture in nomal medium, the BAEC were grown to confluence and their permeability was increased. These findings suggest that B16-CM increased the endothelial permeability irreversibly wi … More thout the decrease of F-actin, and that soluble factor (s) which were secreted from the tumor cells participate in the construction of the hyperpermeable structure of tumor vessels in vivo.Rat KMT-17 fibrosarcoma-derived endothelial cells were isolated by Percoll gradient centrifugation with an attaching-speed separation technique, and their properties in culture were examined. The primary cultured tumor-derived endothelial cells (TEC) showed angiotensin-converting enzyme activity, positivity for Factor VIII-related antigen staining, and typical capillary-like formation on Matrigel. The primary cultured TEC monolayr showed greater permeability than normal tissue-derived endothelial cell (aorta, vena cava and epididymal fat capillary) monolayrs on FITC-dextran diffusion (molecular weight 70,000). Leukocyte adhesion to TEC was reduced compared to that to fat-derived capillary endothelial cells. These characteristics resembled those of tumor vascular endothelium, and were observed both in the primary and first-passage cell cultures, but not in the fourth-passage cell cultures. Our findings indicate that primary or subcultured TEC are applicable for studies of the physiological characteristics of tumor endothelial cells. Less