产品简介: vEPT 细胞系来源于 1989 年,由 M. Romero 和 U. Hopfer 从一只正常 2 公斤雄性新西兰兔肾脏的浅表皮层切片显微解剖近端小管(S1 段)的原代培养物。这些细胞最初是在涂有聚-L-赖氨酸的多孔无机过滤器上生长的。目前,牛I型胶原蛋白被用作基质。经过几次传代后,将细胞与辐照的成纤维细胞共培养,产生编码 SV40、大 T 抗原和 G418 抗性的重组逆转录病毒。然而,SV40 T 抗原表达对于永生化不是必需的,因为在 vEPT 细胞系中既未检测到 SV40 T 抗原,也未检测到 G418 耐药性。这些细胞显示出上皮细胞的特性,形成具有顶端微绒毛、紧密连接和基底外侧质膜卷积的汇合单层。它们保留了近端肾小管和受体的电解质转运特性以及血管紧张素 II 的信号传导机制。它们表达细胞角蛋白 8、10+11 和 19。 产品属性: 产品分类:动物细胞 生物:Oryctolagus cuniculus,兔子 细胞类型:上皮成纤维细胞 细胞形态:上皮细胞样 组织:肾;近端小管 疾病:正常 应用:3D细胞培养 产品格式:冷冻 储存条件:液氮 培养物检查 定期仔细观察培养物的形态和活力。检查容器中的介质是否存在微生物污染的宏观证据。这包括不寻常的pH值变化(由酚红变为黄色或紫色)、浊度或颗粒。此外,寻找漂浮在介质-空气界面的小真菌菌落。具体检查血管边缘周围,因为这些边缘可能不容易通过显微镜看到。 用低倍镜(40×)的倒置显微镜检查培养基是否有微生物污染和细胞形态的证据。细菌污染会在细胞之间的空间内表现为微小的、闪闪发光的黑点。酵母污染会表现为圆形或出芽的颗粒,而真菌会有细丝状菌丝。对于在烧瓶中生长的非粘附细胞,如杂交瘤,这是直接在显微镜上观察烧瓶的简单问题。对于在旋转瓶或生物反应器中生长的细胞,需要提取细胞悬浮液样本,并将其装载到显微镜载玻片或血细胞仪中进行观察。 Examination of cultures Observe the morphology and viability of cultures regularly and carefully. Examine the medium in the vessel for macroscopic evidence of microbial contamination. This includes unusual pH shifts (yellow or purple color from the phenol red), turbidity, or particles. Also, look for small fungal colonies that float at the medium-air interface. Specifically check around the edges of the vessel as these may not be readily visible through the microscope. With an inverted microscope at low power (40×), check the medium for evidence of microbial contamination and the morphology of the cells. Bacterial contamination will appear as small, shimmering black dots within the spaces between the cells. Yeast contamination will appear as rounded or budding particles, while fungi will have thin filamentous mycelia. For nonadherent cells grown in flasks, such as hybridomas, this is a simple matter of viewing the flask directly on the microscope. For cells grown in spinner flasks or bioreactors, a sample of the cell suspension will need to be withdrawn and loaded into a microscope slide or hemocytometer for observation.