Characterization of two blood–brain barrier mimicking cell lines: Distribution of lectin-binding sites and perspectives for drug delivery
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- 作者：V.E. Plattner ; B. Germann ; W. Neuhaus ; C.R. Noe ; F. Gabor ; M. Wirth
- 关键词：Blood– ; brain barrier ; ECV304 ; PBMEC/C1– ; 2 ; Lectins ; Drug targeting
- 刊名：International Journal of Pharmaceutics
- 出版年：2010
- 出版时间：15 March 2010
- 年：2010
- 卷：387
- 期：1-2
- 页码：34-41
- 全文大小：604 K
文摘
In the present study plant lectins with distinct sugar specificities were applied to two blood–brain barrier (BBB) mimicking cell lines, namely human ECV304 and porcine brain microvascular endothelial cells PBMEC/C1–2 in order to elucidate their glycosylation pattern and to evaluate the lectin-cell interaction for lectin-mediated targeting. The bioadhesive properties of fluorescein-labeled lectins were investigated with monolayers as well as single cells using fluorimetry and flow cytometry, followed by confirmation of the specificity of binding. For PBMEC/C1–2 layers highest binding capacity was found for wheat germ agglutinin (WGA), followed by Dolichus biflorus agglutinin (DBA) whereas single cell experiments revealed a predominance of DBA only. Analyzing ECV304 monolayers and single cells, WGA yielded the strongest interaction without any changes during cultivation. The binding capacities of the other lectins increased significantly during differentiation. As similar results to primary cells and brain sections were observed, both cell lines seem to be suitable as models for lectin-interaction studies. Thus, an additional focus was set on the mechanisms involved in uptake and intracellular fate of selected lectins. Cytoinvasion studies were performed with WGA for human ECV304 cells and WGA as well as DBA for PBMEC/C1–2 cells. For both lectins, the association rate to the cells was dependent on temperature which indicated cellular uptake.