bims-orenst 2021-09-26 papers

Issue of 2021–09–26
five papers selected by
Joram Mooiweer, University of Groningen

Front Pharmacol. 2021 ;12 707891

A Synthetic Hydrogel, VitroGel® ORGANOID-3, Improves Immune Cell-Epithelial Interactions in a Tissue Chip Co-Culture Model of Human Gastric Organoids and Dendritic Cells.

Michelle D Cherne, Barkan Sidar, T Andrew Sebrell, Humberto S Sanchez, Kody Heaton, Francis J Kassama, Mandi M Roe, Andrew B Gentry, Connie B Chang, Seth T Walk, Mark Jutila, James N Wilking, Diane Bimczok.

Immunosurveillance of the gastrointestinal epithelium by mononuclear phagocytes (MNPs) is essential for maintaining gut health. However, studying the complex interplay between the human gastrointestinal epithelium and MNPs such as dendritic cells (DCs) is difficult, since traditional cell culture systems lack complexity, and animal models may not adequately represent human tissues. Microphysiological systems, or tissue chips, are an attractive alternative for these investigations, because they model functional features of specific tissues or organs using microscale culture platforms that recreate physiological tissue microenvironments. However, successful integration of multiple of tissue types on a tissue chip platform to reproduce physiological cell-cell interactions remains a challenge. We previously developed a tissue chip system, the gut organoid flow chip (GOFlowChip), for long term culture of 3-D pluripotent stem cell-derived human intestinal organoids. Here, we optimized the GOFlowChip platform to build a complex microphysiological immune-cell-epithelial cell co-culture model in order to study DC-epithelial interactions in human stomach. We first tested different tubing materials and chip configurations to optimize DC loading onto the GOFlowChip and demonstrated that DC culture on the GOFlowChip for up to 20 h did not impact DC activation status or viability. However, Transwell chemotaxis assays and live confocal imaging revealed that Matrigel, the extracellular matrix (ECM) material commonly used for organoid culture, prevented DC migration towards the organoids and the establishment of direct MNP-epithelial contacts. Therefore, we next evaluated DC chemotaxis through alternative ECM materials including Matrigel-collagen mixtures and synthetic hydrogels. A polysaccharide-based synthetic hydrogel, VitroGel®-ORGANOID-3 (V-ORG-3), enabled significantly increased DC chemotaxis through the matrix, supported organoid survival and growth, and did not significantly alter DC activation or viability. On the GOFlowChip, DCs that were flowed into the chip migrated rapidly through the V-ORG matrix and reached organoids embedded deep within the chip, with increased interactions between DCs and gastric organoids. The successful integration of DCs and V-ORG-3 embedded gastric organoids into the GOFlowChip platform now permits real-time imaging of MNP-epithelial interactions and other investigations of the complex interplay between gastrointestinal MNPs and epithelial cells in their response to pathogens, candidate drugs and mucosal vaccines.

Keywords: chemotaxis; dendritic cell; gastric organoid; hydrogel; matrigel; microphysiological system; mononuclear phagocyte

DOI: https://doi.org/10.3389/fphar.2021.707891

Biomicrofluidics. 2021 Sep;15(5): 054102

Microfluidic model of monocyte extravasation reveals the role of hemodynamics and subendothelial matrix mechanics in regulating endothelial integrity.

Sandra Pérez-Rodríguez, Stephanie A Huang, Carlos Borau, José Manuel García-Aznar, William J Polacheck.

Extravasation of circulating cells is an essential process that governs tissue inflammation and the body's response to pathogenic infection. To initiate anti-inflammatory and phagocytic functions within tissues, immune cells must cross the vascular endothelial barrier from the vessel lumen to the subluminal extracellular matrix. In this work, we present a microfluidic approach that enables the recreation of a three-dimensional, perfused endothelial vessel formed by human endothelial cells embedded within a collagen-rich matrix. Monocytes are introduced into the vessel perfusate, and we investigate the role of luminal flow and collagen concentration on extravasation. In vessels conditioned with the flow, increased monocyte adhesion to the vascular wall was observed, though fewer monocytes extravasated to the collagen hydrogel. Our results suggest that the lower rates of extravasation are due to the increased vessel integrity and reduced permeability of the endothelial monolayer. We further demonstrate that vascular permeability is a function of collagen hydrogel mass concentration, with increased collagen concentrations leading to elevated vascular permeability and increased extravasation. Collectively, our results demonstrate that extravasation of monocytes is highly regulated by the structural integrity of the endothelial monolayer. The microfluidic approach developed here allows for the dissection of the relative contributions of these cues to further understand the key governing processes that regulate circulating cell extravasation and inflammation.

DOI: https://doi.org/10.1063/5.0061997

Toxicology. 2021 Sep 17. pii: S0300-483X(21)00277-8. [Epub ahead of print] 152954

Prediction of Hepatic Drug Clearance with a Human Microfluidic Four-Cell Liver Acinus MicroPhysiology System.

Courtney Sakolish, Yu-Syuan Luo, Alan Valdiviezo, Lawrence A Vernetti, Ivan Rusyn, Weihsueh A Chiu.

Predicting human hepatic clearance remains a fundamental challenge in both pharmaceutical drug development and toxicological assessments of environmental chemicals, with concerns about both accuracy and precision of in vitro-derived estimates. Suggested sources of these issues have included differences in experimental protocols, differences in cell sourcing, and use of a single cell type, liver parenchymal cells (hepatocytes). Here we investigate the ability of human microfluidic four-cell liver acinus microphysiology system (LAMPS) to make predictions as to hepatic clearance for seven representative compounds: Caffeine, Pioglitazone, Rosiglitazone, Terfenadine, Tolcapone, Troglitazone, and Trovafloxacin. The model, whose reproducibility was recently confirmed in an inter-lab comparison, was constructed using primary human hepatocytes or human induced pluripotent stem cell (iPSC)-derived hepatocytes and 3 human cell lines for the endothelial, Kupffer and stellate cells. We calculated hepatic clearance estimates derived from experiments using LAMPS or traditional 2D cultures and compared the outcomes with both in vivo human clinical study-derived and in vitro human hepatocyte suspension culture-derived values reported in the literature. We found that, compared to in vivo clinically-derived values, the LAMPS model with iPSC-derived hepatocytes had higher precision as compared to primary cells in suspension or 2D culture, but, consistent with previous studies in other microphysiological systems, tended to underestimate in vivo clearance. Overall, these results suggest that use of LAMPS and iPSC-derived hepatocytes together with an empirical scaling factor warrants additional study with a larger set of compounds, as it has the potential to provide more accurate and precise estimates of hepatic clearance.

Keywords: hepatic clearance; in vitro; in vivo; microphysiological systems; pharmacokinetics; toxicokinetics

DOI: https://doi.org/10.1016/j.tox.2021.152954

Acta Biomater. 2021 Sep 18. pii: S1742-7061(21)00610-3. [Epub ahead of print]

Engineering a 3D Human Intracranial Aneurysm Model using Liquid-Assisted Injection Molding and Tuned Hydrogels.

Kar Wey Yong, Mohsen Janmaleki, Mohammad Pachenari, Alim P Mitha, Amir Sanati-Nezhad, Arindom Sen.

Physiologically relevant intracranial aneurysm (IA) models are crucially required to facilitate testing treatment options for IA. Herein, we report the development of a new in vitro tissue-engineered platform, which recapitulates the microenvironment, structure, and cellular complexity of native human IA. A new modified liquid-assisted injection molding technique was developed to fabricate a three-dimensional hollow IA model with clinically relevant IA dimensions within a mechanically tuned Gelatin Methacryloyl (GelMA) hydrogel. An endothelium lining was created inside the IA model by culturing human umbilical vein endothelial cells over pre-cultured human brain vascular smooth muscle cells. These cellularized IA models were subjected to medium perfusion at flow rates between 6.3 and 15.75 mL/min for inducing biomimetic vessel wall shear stress (10 - 25 dyn/cm2) to the cells for ten days. Both cell types maintained their secretome profiles and showed more than 96% viability, demonstrating the biocompatibility of the hydrogel during perfusion cell culture at such flow rates. Based on the characterized viscoelastic properties of the GelMA hydrogel and with the aid of a fluid-structure interaction model, the capability of the IA model in predicting the response of the IA to different fluid flow profiles was mathematically shown. With physiologically relevant behavior, our developed in vitro human IA model could allow researchers to better understand the pathophysiology and treatment of IA. STATEMENT OF SIGNIFICANCE: • A three-dimensional intracranial aneurysm (IA) tissue model recapitulating the microenvironment, structure, and cellular complexity of native human IA was developed. • An endothelium lining was created inside the IA model over pre-cultured human brain vascular smooth muscle cells over at least 10-day successful culture. • The cells maintained their secretome profiles, demonstrating the biocompatibility of hydrogel during a long-term perfusion cell culture. • The IA model showed its capability in predicting the response of IA to different fluid flow profiles. • The cells in the vessel region behaved differently from cells in the aneurysm region due to alteration in hemodynamic shear stress. • The IA model could allow researchers to better understand the pathophysiology and treatment options of IA.

Keywords: balloon catheter; biomaterial; biomimetic tissue; in vitro hydrogel model; intracranial aneurysm; modified liquid-assisted injection molding

DOI: https://doi.org/10.1016/j.actbio.2021.09.022

Macromol Biosci. 2021 Sep 24. e2100327

Bioprinting of Human Liver-derived Epithelial Organoids for Toxicity Studies.

Manon C Bouwmeester, Paulina Nunez Bernal, Loes A Oosterhoff, Monique van Wolferen, Vivian Lehmann, Monique Vermaas, Maj-Britt Buchholz, Quentin Peiffer, Jos Malda, Luc J W van der Laan, Nynke I Kramer, Kerstin Schneeberger, Riccardo Levato, Bart Spee.

There is a need for long-lived hepatic in vitromodels to better predict drug induced liver injury (DILI). Human liver-derived epithelial organoids are a promising cell source for advanced in vitromodels. Here, organoid technology is combined with biofabrication techniques, which holds great potential for the design of in vitromodels with complex and customizable architectures. In the present study, porous constructs with human hepatocyte-like cells derived from organoids weregenerated using extrusion-based printing technology. Cell viability of bioprinted organoids remained stable for up to ten days (88 - 107% cell viability compared to the day of printing). The expression of hepatic marker G6PC, transporters BSEP and ABCG2 and phase I enzyme CYP3A4 increased compared to undifferentiated controls (expansion condition), and wascomparable to non-printed controls. Exposure to acetaminophen, a well-known hepatotoxic compound, decreased cell viability of bioprinted liver organoids to 21-51% (p<0.05) compared to the start of exposure and elevated levels of damage marker miR-122 wereobserved in the culture medium, indicating the potential use of the bioprinted constructs for toxicity testing. Weshowed that human liver-derived epithelial organoids can be combined with a biofabrication approach, thereby paving the way to create perfusable, complex constructs which can be used as toxicology- and disease-models. This article is protected by copyright. All rights reserved.

Keywords: drug induced liver injury; extrusion-based bioprinting; in vitro modelling; organoids

DOI: https://doi.org/10.1002/mabi.202100327