Decellularization of precision-cut kidney slices-application of physical and chemical methods.

The extracellular matrix (ECM) obtained by decellularization provides scaffolds with the natural complex architecture and biochemical composition of the target organ. Whole kidney decellularization by perfusion uses the vasculature to remove cells leaving a scaffold that can be recellularized with patient-specific cells. However, decellularization and recellularization are highly complex processes that require intensive optimization of various parameters. In pursuit of this, a huge number of animals must be sacrificed. Therefore, we used precision-cut kidney slices (PCKS) as a source of natural scaffolds, which were decellularized by immersion in chemical reagents allowing the examination of more parameters with less animals. However, chemical reagents have a damaging effect on the structure and components of the ECM. Therefore, this study aimed at investigating the effects of physical treatment methods on the effectiveness of PCKS decellularization by immersion in chemical reagents (CHEM). PCKS were treated physically before or during immersion in chemicals (CHEM) with high hydrostatic pressure (HHP), freezing-thawing cycles (FTC) or in an ultrasonic bath system (UBS). Biochemical and DNA quantification as well as structural evaluation with conventional histology and scanning electron microscopy (SEM) were performed. Compared to decellularization by CHEM alone, FTC treatment prior to CHEM was the most effective in reducing DNA while also preserving glycosaminoglycan (GAG) content. Moreover, while UBS resulted in a comparable reduction of DNA, it was the least effective in retaining GAGs. In contrast, despite the pretreatment with HHP with pressures up to 200 MPa, it was the least effective in DNA removal. Histological scoring showed that HHP scaffolds received the best score followed by UBS, FTC and CHEM scaffolds. However further analysis with SEM demonstrated a higher deterioration of the ultrastructure in UBS scaffolds. Altogether, pretreatment with FTC prior to CHEM resulted in a better balance between DNA removal and structural preservation.

Operational locality in global theories.

Within a global physical theory, a notion of locality allows us to find and justify information-processing primitives, like non-signalling between distant agents. Here, we propose exploring the opposite direction: to take agents as the basic building blocks through which we test a physical theory, and recover operational notions of locality from signalling conditions. First, we introduce an operational model for the effective state spaces of individual agents, as well as the range of their actions. We then formulate natural secrecy conditions between agents and identify the aspects of locality relevant for signalling. We discuss the possibility of taking commutation of transformations as a primitive of physical theories, as well as applications to quantum theory and generalized probability frameworks. This 'it from bit' approach establishes an operational connection between local actions and local observations, and gives a global interpretation to concepts like discarding a subsystem or composing local functions.This article is part of a discussion meeting issue 'Foundations of quantum mechanics and their impact on contemporary society'.