Real-time quantitative imaging is becoming highly desirable to study nondestructively the biological behavior of three dimensional cell culture systems. In this work, we investigate the feasibility of quantitative imaging/monitoring of 3D cell culture processes via Electrical Impedance Tomography (EIT), which is capable of generating conductivity images in a non-destructive manner with high temporal resolution. To this end, a planar miniature EIT sensor amenable to standard cell culture format is designed and a 3D forward model for the sensor is developed for 3D imaging. Furthermore, a novel 3D-Laplacian and sparsity joint regularization algorithm is proposed for enhanced 3D image reconstruction. Simulation phantoms with spheres at various vertical and horizontal positions were imaged for 3D performance evaluation. In addition, experiments on human breast cancer cell spheroid and a triangular breast cancer cell pellet were carried out for experimental verification. The results have shown that stable measurement on high conductive cell culture medium and significant improvement of image quality based on the proposed regularization method are achieved. It demonstrates the feasibility of using the miniature EIT sensor and 3D image reconstruction algorithm to visualize 3D cell cultures such as spheroids or artificial tissues and organs. The established work would expedite real-time quantitative imaging of 3D cell culture for assessment of cellular dynamics.
Therapies based on regenerative techniques have the potential to radically improve healthcare in the coming years. As a result, there is an emerging need for non-destructive and label-free technologies to assess the quality of engineered tissues and cell-based products prior to their use in the clinic. In parallel, the emerging regenerative medicine industry that aims to produce stem cells and their progeny on a large scale will benefit from moving away from existing destructive biochemical assays towards data-driven automation and control at the industrial scale. Impedance-based cellular assays (IBCA) have emerged as an alternative approach to study stem-cell properties and cumulative studies, reviewed here, have shown their potential to monitor stem-cell renewal, differentiation and maturation. They offer a novel method to non-destructively assess and quality-control stem-cell cultures. In addition, when combined with disease models they provide complementary insights as label-free phenotypic assays. IBCA provide quantitative and very sensitive results that can easily be automated and up-scaled in multi-well format. When facing the emerging challenge of real-time monitoring of three-dimensional cell culture dielectric spectroscopy and electrical impedance tomography represent viable alternatives to two-dimensional impedance sensing.This article is part of the theme issue 'Designer human tissue: coming to a lab near you'.
Dysfunction of cell-cell tight junction (TJ) adhesions is a major feature in the pathogenesis of various diseases. Liver TJs preserve cellular polarity by delimiting functional bile-canalicular structures, forming the blood-biliary barrier. In acetaminophen-hepatotoxicity, the mechanism by which tissue cohesion and polarity are affected remains unclear. Here, we demonstrate that acetaminophen, even at low-dose, disrupts the integrity of TJ and cell-matrix adhesions, with indicators of cellular stress with liver injury in the human hepatic HepaRG cell line, and primary hepatocytes. In mouse liver, at human-equivalence (therapeutic) doses, dose-dependent loss of intercellular hepatic TJ-associated ZO-1 protein expression was evident with progressive clinical signs of liver injury. Temporal, dose-dependent and specific disruption of the TJ-associated ZO-1 and cytoskeletal-F-actin proteins, correlated with modulation of hepatic ultrastructure. Real-time impedance biosensing verified in vitro early, dose-dependent quantitative decreases in TJ and cell-substrate adhesions. Whereas treatment with NAPQI, the reactive metabolite of acetaminophen, or the PKCα-activator and TJ-disruptor phorbol-12-myristate-13-acetate, similarly reduced TJ integrity, which may implicate oxidative stress and the PKC pathway in TJ destabilization. These findings are relevant to the clinical presentation of acetaminophen-hepatotoxicity and may inform future mechanistic studies to identify specific molecular targets and pathways that may be altered in acetaminophen-induced hepatic depolarization.
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