A novel organotypic tauopathy model on a new microcavity chip for bioelectronic label-free and real time monitoring.

Autor: Krinke D; Centre for Biotechnology and Biomedicine (BBZ), University of Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Deutscher Platz 5, 04103 Leipzig, Germany., Jahnke HG, Mack TG, Hirche A, Striggow F, Robitzki AA
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2010 Sep 15; Vol. 26 (1), pp. 162-8. Date of Electronic Publication: 2010 Jun 09.
DOI: 10.1016/j.bios.2010.06.002
Abstrakt: Herewith we developed a novel 3D in vitro Alzheimer's disease (AD) model, based on the human neuroblastoma cell line SH-SY5Y, which is well differentiated without the application of any agents. Furthermore AD-like pathological neurodegeneration can be induced by okadaic acid (OA) mediated hyperphosphorylation of the microtubule associated protein tau. Moreover, we established stable "rapid tauopathy cell lines" expressing additional EGFP-fused (enhanced green fluorescent protein) wildtype or a pathology-promoting mutant tau variant (P301L) by lentiviral transduction. For the sensitive and feasible quantitative detection of pathological effects on neuronal 3D-cultures by electrochemical impedance spectroscopy (EIS) we optimized and redesigned a microcavity array (MCA). The cellular contribution to impedance could be increased by the factor of 2.5 and the variance decreased by 40%. Using our optimized MCA and impedance measurement setup we were able to detect quantitatively an OA concentration- and time-dependent decrease of the impedance in 3D SH-SY5Y cultures. Moreover, we were able to detect and quantify distinct, AD-related effects triggered by tau-mutant (P301L) expression and hyperphosphorylation in our organotypic 3D-cultures with the help of impedance spectroscopy.
(Copyright 2010 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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