Induced Tauopathy in a Novel 3D-Culture Model Mediates Neurodegenerative Processes: A Real-Time Study on Biochips

Abstract

Tauopathies including Alzheimer’s disease represent one of the major health problems of aging population worldwide. Therefore, a better understanding of tau-dependent pathologies and consequently, tau-related intervention strategies is highly demanded. In recent years, several tau-focused therapies have been proposed with the aim to stop disease progression. However, to develop efficient active pharmaceutical ingredients for the broad treatment of Alzheimer’s disease patients, further improvements are necessary for understanding the detailed neurodegenerative processes as well as the mechanism and side effects of potential active pharmaceutical ingredients (API) in the neuronal system. In this context, there is a lack of suitable complex in vitro cell culture models recapitulating major aspects of taupathological degenerative processes in sufficient time and reproducible manner.Herewith, we describe a novel 3D SH-SY5Y cell-based, tauopathy model that shows advanced characteristics of matured neurons in comparison to monolayer cultures without the need of artificial differentiation promoting agents. Moreover, the recombinant expression of a novel highly pathologic fourfold mutated human tau variant lead to a fast and emphasized degeneration of neuritic processes. The neurodegenerative effects could be analyzed in real time and with high sensitivity using our unique microcavity array-based impedance spectroscopy measurement system. We were able to quantify a time- and concentration-dependent relative impedance decrease when Alzheimer’s disease-like tau pathology was induced in the neuronal 3D cell culture model. In combination with the collected optical information, the degenerative processes within each 3D-culture could be monitored and analyzed. More strikingly, tau-specific regenerative effects caused by tau-focused active pharmaceutical ingredients could be quantitatively monitored by impedance spectroscopy.Bringing together our novel complex 3D cell culture taupathology model and our microcavity array-based impedimetric measurement system, we provide a powerful tool for the label-free investigation of tau-related pathology processes as well as the high content analysis of potential active pharmaceutical ingredient candidates.

A novel 96-well multielectrode array based impedimetric monitoring platform for comparative drug efficacy analysis on 2D and 3D brain tumor cultures

“…For 3D cultures, the microcavity array based impedimetric measurement is performed manually (Kloss et al, 2008a;Krinke et al, 2010) without the need to keep the spheroids on the chip during the whole experiment. In this way, up to 800 spheroids can be measured by an individual experimenter on a single chip and measurement system per day (Seidel et al, 2012). In contrast, small scale multiwell-electrode arrays (Haas et al, 2010) with adhered 2D cultures are automatically and continuously measured with a single measurement setup, throughout the whole experiment.…”

“…Since 2D cultures have shown to be an expedient but still highly artificial representation of the in vivo situation (Hirschhaeuser et al, 2010), 3D cultures have been introduced to mimic the corresponding in vivo tissues and allow studying of basic mechanisms like cell growth, proliferation and differentiation in a reproducible format (Kunz-Schughart et al, 1998) as well as for testing novel treatment strategies (Hirschhaeuser et al, 2010). In this context, bioelectronic monitoring techniques like impedance spectroscopy in combination with novel microcavity arrays were used to quantify cellular damage and degenerative processes in 3D cultures (Kloss et al, 2008a(Kloss et al, , 2008bKrinke et al, 2010;Seidel et al, 2012). Moreover, quantification of immunotoxin efficacy in tumor spheroids compared to known small molecule reference chemotherapeutics was demonstrated by impedance spectroscopy (Poenick et al, 2014).…”

In Vitro Modeling of Tissue-Specific 3D Microenvironments and Possibile Application to Pediatric Cancer Research

“…In 3D conditions, Matrigel TM invasion chamber assay is one of the most popular method [299][300][301][302][303][304][305], together with the invasion assays through 3D collagen gels [306]. Thanks to these studies, the effects of ionizing radiation [307], differentiating agents [299,301], antitumoral drugs/molecules [303][304][305] or the contribution of specific oncogenes [300,302] on invasiveness have been explored.…”

“…Neuroblastoma 3D culture embedded in soft matrices, like agarose, Matrigel TM or collagen, is widely used by scientists [299][300][301][302][303][304][305]308]. Valero and coworkers [309], have studied the impact of different soft matrices on neuronal differentiation of the Neuro2a neuroblastoma cell line (N2a), evaluated with a novel impedimetric biosensor.…”

Experimental Models for Identifying Modifiers of Polyglutamine-Induced Aggregation and Neurodegeneration

“…For example, 3D hydrogel tissue constructs formed through extracellular matrix remodeling can measure contractile force, which does not occur in twodimensional culture [70]. Similarly, 3D spheroid cultures derived from SH-SY5Y neuroblastoma cells expressing tau variants have been used as a model of tauopathy [71].…”