Centre for Biotechnology and Biomedicine (BBZ), University of Leipzig, Division of Molecular Biological-Biochemical Processing Technology, Leipzig, Germany.
PLoS One. 2012;7(11):e49150. doi: 10.1371/journal.pone.0049150. Epub 2012 Nov 7.
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.
包括阿尔茨海默病在内的 tau 病是全球老龄化人口的主要健康问题之一。因此，人们强烈需要更好地了解 tau 依赖性病理，并因此，tau 相关的干预策略。近年来，已经提出了几种针对 tau 的治疗方法，旨在阻止疾病的进展。然而，为了开发针对广泛的阿尔茨海默病患者的高效活性药物成分，需要进一步改进以了解详细的神经退行性过程以及潜在的活性药物成分（API）在神经元系统中的机制和副作用。在这方面，缺乏合适的复杂体外细胞培养模型来充分时间和可重复的方式再现 taupathological 退行性过程的主要方面。在这里，我们描述了一种新颖的 3D SH-SY5Y 细胞基 tau 病模型，与单层培养物相比，该模型显示出成熟神经元的高级特征，而无需使用人工分化促进剂。此外，新型高度病理四倍突变人 tau 变体的重组表达导致神经突过程快速而强调的退化。使用我们独特的基于微腔阵列的阻抗谱测量系统可以实时和高灵敏度地分析神经退行性效应。当在神经元 3D 细胞培养模型中诱导类似于阿尔茨海默病的 tau 病理时，我们能够定量地测量到时间和浓度依赖性的相对阻抗降低。与收集的光学信息相结合，可以监测和分析每个 3D 培养物中的退化过程。更引人注目的是，通过阻抗谱可以定量监测针对 tau 的活性药物成分的特异性再生作用。通过将我们的新型复杂 3D 细胞培养 tau 病模型与我们的基于微腔阵列的阻抗测量系统相结合，我们提供了一种强大的工具，用于无标记研究 tau 相关的病理过程以及潜在的活性药物成分候选物的高内涵分析。
