[Models of glomerular filtration barrier : New developments]. / Modélisation de la barrière de filtration glomérulaire - Nouvelles avancées.

In this article, we present the latest innovations to generate in vitro models of the glomerular filtration barrier. There is currently a growing interest for such model systems that allow to reduce the use of animal models. Methodologies to improve their physiological relevance have taken advantage of the development of induced pluripotent stem cells and of bioengineering, particularly tissue engineering. Here, we first introduce the methods to overcome the limitations of the currently used glomerular cells based on the use of stem cells. The different approaches to obtain podocytes, the most important cells in the glomerulus, are presented. Finally, we emphasize the importance of the glomerular microenvironment in maintaining the glomerular cell phenotype, which can be achieved by co-culturing different glomerular cells, integration of biomaterials mimicking the extracellular matrix and introduction of flows with microfluidics.

TITLE: Modélisation de la barrière de filtration glomérulaire - Nouvelles avancées. ABSTRACT: Nous présentons, dans cette revue, les dernières avancées concernant la modélisation in vitro de la barrière de filtration glomérulaire. Ces systèmes, permettant de réduire l'utilisation des modèles animaux, connaissent un intérêt croissant et bénéficient du développement de nos connaissances des cellules souches et de la bioingénierie. Nous discuterons les limites des modèles cellulaires glomérulaires actuels et nous introduirons les méthodes permettant d'obtenir des cellules glomérulaires à partir des cellules souches. Enfin, nous discuterons de l'importance du microenvironnement dans le maintien du phénotype, quels que soient les systèmes utilisés tels que la co-culture, les biomatériaux ou la microfluidique.

Designer artificial membrane binding proteins to direct stem cells to the myocardium.

We present a new cell membrane modification methodology where the inherent heart tissue homing properties of the infectious bacteria Streptococcus gordonii are transferred to human stem cells. This is achieved via the rational design of a chimeric protein-polymer surfactant cell membrane binding construct, comprising the cardiac fibronectin (Fn) binding domain of the bacterial adhesin protein CshA fused to a supercharged protein. Significantly, the protein-polymer surfactant hybrid spontaneously inserts into the plasma membrane of stem cells without cytotoxicity, instilling the cells with a high affinity for immobilized fibronectin. Moreover, we show that this cell membrane reengineering approach significantly improves retention and homing of stem cells delivered either intracardially or intravenously to the myocardium in a mouse model.

Engineered basement membranes: from in vivo considerations to cell-based assays.

Improvements in the physiological relevance of cell-based assays have been enabled by the development of various interdisciplinary methods. However, due to their complexity, in vivo structures such as basement membranes (BMs), which regulate the phenotype of adherent cells, are still difficult to mimic in vitro. The reconstruction of a physiologically relevant BM is crucially important to develop cell-based assays with the capacity for drug screening and disease modelling. Here, we review the biophysical and biochemical properties of BMs in vivo and their interactions with neighbouring cells. We discuss the current methods used to mimic BM functions in cell-based assays according to the type of targeted applications. In doing so, we examine the advantages and limitations of each method as well as exploring approaches to improve the physiological relevance of engineered or cell-derived BMs in vitro.

Three-Dimensional Graphene: A Biocompatible and Biodegradable Scaffold with Enhanced Oxygenation.

Owing to its high porosity, specific surface area and three-dimensional structure, three-dimensional graphene (3D-C) is a promising scaffold material for tissue engineering, regenerative medicine as well as providing a more biologically relevant platform for living organisms in vivo studies. Recently, its differentiation effects on cells growth and anti-inflammation properties have also been demonstrated. Here, we report a complete study of 3D-C as a fully adequate scaffold for tissue engineering and systematically analyze its biocompatibility and biodegradation mechanism. The metabolic activities of liver cells (HepG2 hepatocarcinoma cells) on 3D-C are studied and our findings show that cell growth on 3D-C has high cell viability (> 90%), low lactate production (reduced by 300%) and its porous structure also provides an excellent oxygenation platform. 3D-C is also biodegradable via a 2-step oxidative biodegradation process by first, disruption of domains and lift off of smaller graphitic particles from the surface of the 3D-C and subsequently, the decomposition of these graphitic flakes. In addition, the speed of the biodegradation can be tuned with pretreatment of O2 plasma.

New physiologically-relevant liver tissue model based on hierarchically cocultured primary rat hepatocytes with liver endothelial cells.

To develop an in vitro liver tissue equivalent, hepatocytes should be cocultured with liver non-parenchymal cells to mimic the in vivo physiological microenvironments. In this work, we describe a physiologically-relevant liver tissue model by hierarchically organizing layers of primary rat hepatocytes and human liver sinusoidal endothelial cells (TMNK-1) on an oxygen-permeable polydimethylsiloxane (PDMS) membrane, which facilitates direct oxygenation by diffusion through the membrane. This in vivo-mimicking hierarchical coculture was obtained by simply proceeding the overlay of TMNK-1 cells on the hepatocyte layer re-formed on the collagen immobilized PDMS membranes. The comparison of hepatic functionalities was achieved between coculture and sandwich culture with Matrigel, in the presence and absence of direct oxygenation. A complete double-layered structure of functional liver cells with vertical contact between hepatocytes and TMNK-1 was successfully constructed in the coculture with direct oxygen supply and was well-maintained for 14 days. The hepatocytes in this hierarchical culture exhibited improved survival, functional bile canaliculi formation, cellular level polarization and maintenance of metabolic activities including Cyp1A1/2 activity and albumin production. By contrast, the two cell populations formed discontinuous monolayers on the same surfaces in the non-oxygen-permeable cultures. These results demonstrate that (i) the direct oxygenation through the PDMS membranes enables very simple formation of a hierarchical structure consisting of a hepatocyte layer and a layer of TMNK-1 and (ii) we may include other non-parenchymal cells in this format easily, which can be widely applicable to other epithelial organs.

Inhibiting protein biofouling using graphene oxide in droplet-based microfluidic microsystems.

Biofouling or adsorption of biomolecules onto surfaces in microfluidic devices limits the type of samples which can be handled. In this paper, we take advantage of the high adsorption capacity of graphene oxide (GO) for proteins as a strategy to limit biofouling, while preserving their activity for droplet-based lab-on-chip applications.

Sliding droplets on superomniphobic zinc oxide nanostructures.

This study reports on liquid-repellency of zinc oxide nanostructures (ZnO NS). The ZnO NS are synthesized by an easy and fast chemical bath deposition technique. Three different nanostructured surfaces consisting of nanorods, flowers, and particles are prepared, depending on the deposition time and the presence of ethanolamine in the reaction mixture. Chemical functionalization of the ZnO NS with 1H,1H,2H,2H-perfluorodecyltrichlorosilane (PFTS) in liquid (PFTS L) and vapor phase (PFTS V) or through octafluorobutane (C(4)F(8)) plasma deposition led to the formation of superomniphobic surfaces. A comprehensive characterization of the wetting properties (static contact angle and contact angle hysteresis) has been performed using liquids composed of deionized water and various concentrations of ethanol (surface tension between 35 and 72.6 mN/m). Depending on the nanostructures morphology, coating nature and liquid employed, high static apparent contact angles θ ≈ 150-160°, and low contact angle hysteresis Δθ ≈ 0° are obtained. The different ZnO NS are characterized using scanning electron microscopy (SEM) and contact angle measurements. The results reported in this work permit preparation of sliding omniphobic surfaces using a simple and low cost technique.