Bernd Alexandra, Ott Melanie, Ishikawa Hiroshi, Schroten Horst, Schwerk Christian, Fricker Gert
Institute of Pharmacy and Molecular Biotechnology, Ruprecht-Karls University Heidelberg, Im Neuenheimer Feld 329, 69120, Heiderlberg, Germany.
Pharm Res. 2015 Sep;32(9):2973-82. doi: 10.1007/s11095-015-1679-1. Epub 2015 May 19.
To characterize the human choroid plexus (CP) papilloma cell line HIBCPP with respect to ABC export protein expression and function in order to evaluate its use as an in vitro model to study carrier-mediated transport processes at the CP.
Expression profiles of ABC transporters were studied by quantitative real-time PCR and Western Blot analysis. Functionality of transporters was investigated by means of uptake experiments and permeation studies carried out on permeable filter systems. In addition, immunohistochemistry served to study localization of ABCC1 and ABCC4.
Both qPCR and Western Blot revealed that ABC transporters known to be expressed in CP are also expressed in HIBCPP cells. Immunohistochemistry confirmed basolateral expression of ABCC1. Functionality of ABCC1, ABCC4, ABCB1 and ABCG2 could be shown in uptake assays.
Altogether, the HIBCPP cells promise to be a functional and relevant in vitro tool to investigate transport processes at the blood-cerebrospinal fluid barrier.
对人脉络丛(CP)乳头状瘤细胞系HIBCPP的ABC转运蛋白表达和功能进行表征,以评估其作为体外模型用于研究CP处载体介导的转运过程的适用性。
通过定量实时PCR和蛋白质免疫印迹分析研究ABC转运蛋白的表达谱。通过在可渗透滤器系统上进行摄取实验和渗透研究来研究转运蛋白的功能。此外,免疫组织化学用于研究ABCC1和ABCC4的定位。
qPCR和蛋白质免疫印迹均显示已知在CP中表达的ABC转运蛋白也在HIBCPP细胞中表达。免疫组织化学证实ABCC1在基底外侧表达。在摄取试验中可显示ABCC1、ABCC4、ABCB1和ABCG2的功能。
总体而言,HIBCPP细胞有望成为研究血脑脊液屏障转运过程的功能性相关体外工具。