bims-orenst 2022-01-30 papers

bims-orenst

Biomed News

on Organs-on-chips and engineered stem cell models

Issue of 2022–01–30
eight papers selected by
Joram Mooiweer, University of Groningen

Comparative study of the transcriptomes of Caco-2 cells cultured under dynamic vs. static conditions following exposure to titanium dioxide and zinc oxide nanomaterials.
Human endothelial cells in high glucose: New clues from culture in 3D microfluidic chips.
Biomimetic Human lung-on-a-chip with Colorful Displaying Microphysiological Breath.
Spatially arranged encapsulation of stem cell spheroids within hydrogels for the regulation of spheroid fusion and cell migration.
The effects of citrus flavonoids and their metabolites on immune-mediated intestinal barrier disruption using an in vitro co-culture model.
Intrinsic blood-brain barrier dysfunction contributes to multiple sclerosis pathogenesis.
Liver Stromal Cells Restrict Macrophage Maturation and Stromal IL-6 Limits the Differentiation of Cirrhosis-linked Macrophages.
Impact of hydrogel stiffness on the induced neural stem cells modulation.

Nanotoxicology. 2022 Jan 25. 1-20

Comparative study of the transcriptomes of Caco-2 cells cultured under dynamic vs. static conditions following exposure to titanium dioxide and zinc oxide nanomaterials.

Kornphimol Kulthong, Guido J E J Hooiveld, Loes P M Duivenvoorde, Ignacio Miro Estruch, Hans Bouwmeester, Meike van der Zande.

  Due to the widespread application of food-relevant inorganic nanomaterials, the gastrointestinal tract is potentially exposed to these materials. Gut-on-chip in vitro systems are proposed for the investigation of compound toxicity as they better recapitulate the in vivo human intestinal environment than static models, due to the added shear stresses associated with the flow of the medium. We aimed to compare cellular responses of intestinal epithelial Caco-2 cells at the gene expression level upon TiO2 (E171) and ZnO (NM110) nanomaterial exposure when cultured under dynamic and conventionally applied static conditions. Whole-genome transcriptome analyses upon exposure of the cells to TiO2 and ZnO nanomaterials revealed differentially expressed genes and related biological processes that were culture condition specific. The total number of differentially expressed genes (p < 0.01) and affected pathways (p < 0.05 and FDR < 0.25) after nanomaterial exposure was higher under dynamic culture conditions than under static conditions for both nanomaterials. The observed increase in nanomaterial-induced responses in the gut-on-chip model indicates that shear stress might be a major factor in cell susceptibility. This is the first report on the application of a gut-on-chip system in which gene expression responses upon TiO2 and ZnO nanomaterial exposure are evaluated and compared to a static system. It extends current knowledge on nanomaterial toxicity assessment and the influence of a dynamic environment on cellular responses. Application of the gut-on-chip system resulted in higher sensitivity of the cells and might thus be an attractive system for use in the toxicological hazard characterization of nanomaterials.

Keywords:  Gut-on-chip; nanotoxicology; titanium dioxide; transcriptomics; zinc oxide

DOI:  https://doi.org/10.1080/17435390.2021.2012609
FASEB J. 2022 Feb;36(2): e22137

Human endothelial cells in high glucose: New clues from culture in 3D microfluidic chips.

Laura Locatelli, Mehdi Inglebert, Roberta Scrimieri, Priti Kumari Sinha, Gian Vincenzo Zuccotti, Paolo Milani, Lionel Bureau, Chaouqi Misbah, Jeanette A M Maier.

  Several studies have demonstrated the role of high glucose in promoting endothelial dysfunction utilizing traditional two-dimensional (2D) culture systems, which, however, do not replicate the complex organization of the endothelium within a vessel constantly exposed to flow. Here we describe the response to high glucose of micro- and macro-vascular human endothelial cells (EC) cultured in biomimetic microchannels fabricated through soft lithography and perfused to generate shear stress. In 3D macrovascular EC exposed to a shear stress of 0.4 Pa respond to high glucose with cytoskeletal remodeling and alterations in cell shape. Under the same experimental conditions, these effects are more pronounced in microvascular cells that show massive cytoskeletal disassembly and apoptosis after culture in high glucose. However, when exposed to a shear stress of 4 Pa, which is physiological in the microvasculature, human dermal microvascular endothelial cells (HDMEC) show alterations of the cytoskeleton but no apoptosis. This result emphasizes the sensitivity of HDMEC to different regimens of flow. No significant variations in the thickness of glycocalyx were detected in both human endothelial cells from the umbilical vein and HDMEC exposed to high glucose in 3D, whereas clear differences emerge between cells cultured in static 2D versus microfluidic channels. We conclude that culture in microfluidic microchannels unveils unique insights into endothelial dysfunction by high glucose.

Keywords:  cytoskeleton; diabetes; endothelium; glucose; microfluidics; microvasculature on a chip

DOI:  https://doi.org/10.1096/fj.202100914R
Adv Mater. 2022 Jan 22. e2108972

Biomimetic Human lung-on-a-chip with Colorful Displaying Microphysiological Breath.

Yujuan Zhu, Lingyu Sun, Yu Wang, Lijun Cai, Zhuohao Zhang, Yixuan Shang, Yuanjin Zhao.

  Lung-on-a-chip models hold great promise for disease modeling and drug screening in the past decade. Herein, inspired by the iridescence phenomenon of soap bubbles, we present a novel biomimetic 3D microphysiological lung-on-a-chip system with breathing visuality. The system with an array of pulmonary alveoli at the physiological scale was constructed and coated with structural color materials. Cyclic deformation was induced by regular airflow, resembling the expansion and contraction of the alveoli during the rhythmic breathing. As the deformation were accompanied with corresponding synchronous shifts in the structural colors, the constructed system offered the self-reporting of cell mechanics and enabled real-time monitoring of the cultivation process. Using this system, we have investigated the dynamic relationships between color atlas and disease symptoms, showing the essential role of mechanical stretch in the phenotypes of idiopathic pulmonary fibrosis. These features make this human lung system ideal in biological study, disease monitoring and drug discovery. This article is protected by copyright. All rights reserved.

Keywords:  idiopathic pulmonary fibrosis; lung-on-a-chip; mechanical visual; microfluidics; structural color

DOI:  https://doi.org/10.1002/adma.202108972
Acta Biomater. 2022 Jan 24. pii: S1742-7061(22)00057-5. [Epub ahead of print]

Spatially arranged encapsulation of stem cell spheroids within hydrogels for the regulation of spheroid fusion and cell migration.

Se-Jeong Kim, Hayeon Byun, Sangmin Lee, Eunhyung Kim, Gyeong Min Lee, Seung Jae Huh, Jinmyoung Joo, Heungsoo Shin.

  Mesenchymal stem cell spheroids have been encapsulated in hydrogels for various applications because spheroids demonstrate higher cell activity than individual cells in suspension. However, there is limited information on the effect of distance between spheroids (inter-spheroid distance) on fusion or migration in a hydrogel. In this study, we developed temperature-responsive hydrogels with surface microwell patterns to culture adipose-derived stem cell (ASC) spheroids and deliver them into a Matrigel for the investigation of the effect of inter-spheroid distance on spheroid behavior. The ASC spheroids were encapsulated successfully in a Matrigel, denoted as sandwich culture, with a specific inter-spheroid distance ranging from 100 to 400 µm. Interestingly, ASCs migrated from the host spheroid and formed a bridge-like structure between spheroids, denoted as a cellular bridge, only when the inter-spheroid distance was 200 µm. Thus, we performed a sandwich culture of human umbilical vein endothelial cells (HUVECs) and ASCs in co-cultured spheroids in the Matrigel to create a homogeneous endothelial cell network in the hydrogel. The HUVECs sprouted through the ASC cellular bridge and directly interacted with the adjacent spheroid when the inter-spheroid distance was 200 µm. Similar results were obtained from an in vivo study. Thus, our study suggests the appropriate inter-spheroid distance for effective spheroid encapsulation in a hydrogel. STATEMENT OF SIGNIFICANCE: Recently, spheroid-based 3D tissue culture techniques such as spheroid encapsulation or 3D printing are being intensively investigated for various purposes. However, there is limited research regarding the effect of the inter-spheroid distance on spheroid communication. Here, we demonstrate a spatially arranged spheroid encapsulation method within a Matrigel by using a temperature-responsive hydrogel. Human adipose-derived stem cell spheroids are encapsulated with a precisely controlled inter-spheroid distance from 100 to 400 µm and show different tendencies in cell migration and spheroid fusion. Our results suggest that the inter-spheroid distance affects spheroid communication, and thus, the inter-spheroid distance needs to be considered carefully according to the purpose.

Keywords:  3D cell culture; Spheroid; encapsulation; micropattern; spheroid-spheroid interaction

DOI:  https://doi.org/10.1016/j.actbio.2022.01.047
Br J Nutr. 2022 Jan 28. 1-24

The effects of citrus flavonoids and their metabolites on immune-mediated intestinal barrier disruption using an in vitro co-culture model.

Y Stevens, T de Bie, I Pinheiro, M Elizalde, A Masclee, D Jonkers.

  Hesperidin and naringin are citrus flavonoids with known anti-oxidative and anti-inflammatory properties. Evidence from previous studies indicates that both these compounds and the metabolites that are formed during intestinal metabolism are able to exert beneficial effects on intestinal barrier function and inflammation. However, so far, studies investigating the relative contributions of the various compounds are lacking. Therefore, we assessed the effect of citrus flavonoids and their intestinal metabolites on immune-mediated barrier disruption in an in vitro co-culture model. Caco-2 cell monolayers were placed in co-culture with PMA-stimulated THP-1-BlueTM NF-κB cells for 30 hours. At baseline, the citrus flavonoids and their metabolites were added to the apical compartment (50 or 100 µM per compound). After 24 hours, THP-1 cells were incubated with LPS in the basolateral compartment for 6 hours. Incubation with citrus flavonoids and their metabolites did not induce changes in transepithelial electrical resistance, FITC-D4 permeation or gene expression of barrier related genes for any of the compounds tested. After LPS stimulation, NF-κB activity was significantly inhibited by all compounds (100 µM) except for one metabolite (all p ≤ 0.03). LPS-induced production of the cytokines IL-8, TNF-α and IL-6 was inhibited by most compounds (all p < 0.05). However, levels of IL-1β were increased, which may contribute to the lack of an improved barrier effect. Overall, these results suggest that citrus flavonoids may decrease intestinal inflammation via reduction of NF-κB activity and that the parent compounds and their metabolites formed during intestinal metabolism are able to exert comparable effects.

Keywords:  Citrus flavonoids; co-culture model; immune-mediated barrier disruption; intestinal barrier; intestinal inflammation

DOI:  https://doi.org/10.1017/S0007114521004797
Brain. 2022 Jan 27. pii: awac019. [Epub ahead of print]

Intrinsic blood-brain barrier dysfunction contributes to multiple sclerosis pathogenesis.

Hideaki Nishihara, Sylvain Perriot, Benjamin D Gastfriend, Marel Steinfort, Celine Cibien, Sasha Soldati, Kinya Matsuo, Sarah Guimbal, Amandine Mathias, Sean P Palecek, Eric V Shusta, Renaud Du Pasquier, Britta Engelhardt.

  Blood-brain barrier (BBB) breakdown and immune cell infiltration into the central nervous system (CNS) are early hallmarks of multiple sclerosis (MS). The mechanisms leading to BBB dysfunction are incompletely understood and generally thought to be a consequence of neuroinflammation. Here, we have challenged this view and asked if intrinsic alterations in the BBB of MS patients contribute to MS pathogenesis. To this end, we made use of human induced pluripotent stem cells (hiPSCs) derived from healthy controls (HC) and MS patients and differentiated them into brain microvascular endothelial cell (BMEC)-like cells as in vitro model of the BBB. MS-derived BMEC-like cells showed impaired junctional integrity, barrier properties and efflux pump activity when compared to HC. Also, MS-derived BMEC-like cells displayed an inflammatory phenotype with increased adhesion molecule expression and immune cell interactions. Activation of Wnt/β-catenin signaling in MS-derived endothelial progenitor cells enhanced barrier characteristics and reduced the inflammatory phenotype. Our study provides evidence for an intrinsic impairment of BBB function in MS patients that can be modeled in vitro. Human iPSC-derived BMEC-like cells are thus suitable to explore the molecular underpinnings of BBB dysfunction in MS and will assist in the identification of potential novel therapeutic targets for BBB stabilization.

Keywords:  blood-brain barrier; human induced pluripotent stem cells; immune cell migration; multiple sclerosis; permeability

DOI:  https://doi.org/10.1093/brain/awac019
J Hepatol. 2022 Jan 21. pii: S0168-8278(22)00007-1. [Epub ahead of print]

Liver Stromal Cells Restrict Macrophage Maturation and Stromal IL-6 Limits the Differentiation of Cirrhosis-linked Macrophages.

Erica Buonomo, Shenglin Mei, Samantha Guinn, Isabelle Leo, Michael Peluso, Mei-An Nolan, Frank Schildberg, Lei Zhao, Christine Lian, Shuyun Xu, Joseph Misdraji, Peter Kharchenko, Arlene Sharpe.

   BACKGROUND AND AIMS: Myeloid cells are key regulators of cirrhosis, a major cause of mortality worldwide. Because stromal cells can modulate the functionality of myeloid cells in vitro, targeting stromal-myeloid interactions has become an attractive potential therapeutic strategy. We aimed to investigate how human liver stromal cells impact myeloid cell properties and to understand the utility of stromal-myeloid coculture systems to study these interactions in the context of cirrhosis.
METHODS: Single cell RNA sequencing analyses of noncirrhotic (n=7) and cirrhotic (n=5) human liver tissue was correlated to the bulk RNA sequencing results of in vitro cocultured human CD14+ and primary liver stromal cells. Complimentary mechanistic experiments and flow cytometric analysis were performed on human liver stromal-myeloid coculture systems.
RESULTS: We find that stromal-myeloid coculture reduces the frequency CD14+ cell subsets transcriptionally similar to liver macrophages, showing that stromal cells inhibit the maturation of monocytes into macrophages. Stromal cells also influenced in vitro macrophage differentiation by skewing away from cirrhosis-linked CD9+ scar-associated macrophage-like cells and towards CD163+ Kupffer Cell-like macrophages. We identify IL-6 production as a mechanism by which stromal cells limit CD9+ macrophage differentiation and find that local IL-6 levels are decreased in early-stage human liver disease compared to healthy liver tissue, suggesting a protective role for local IL-6 in healthy liver.
CONCLUSIONS: Our work reveals an unanticipated role for liver stromal cells to impede the maturation and alter the differentiation of macrophages and prompts investigation into the role of local IL-6 production in the pathogenesis of liver disease. These studies provide a framework for investigating macrophage-stromal interactions during cirrhosis.
LAY SUMMARY: The impact of human liver stromal cells on myeloid cell maturation and differentiation in liver disease is incompletely understood. In this study, we present a mechanistic analysis using a primary in vitro human liver stromal-myeloid coculture system that is translated to liver disease using scRNA-seq analysis of cirrhotic and noncirrhotic human liver tissue. Our work supports a role for stromal cell contact in restricting macrophage maturation and stromal-secreted IL-6 in limiting the differentiation of a cirrhotic macrophage subset.

Keywords:  IL-6; Liver cirrhosis; human; macrophage differentiation; macrophage maturation; myeloid cells; single cell RNA-seq; stromal cells

DOI:  https://doi.org/10.1016/j.jhep.2021.12.036
Ann Transl Med. 2021 Dec;9(24): 1784

Impact of hydrogel stiffness on the induced neural stem cells modulation.

Yuyan Liang, Sijie Li, Yujia Li, Mo Li, Xiaohong Sun, Jing An, Qunyuan Xu, Zhiguo Chen, Ying Wang.

   Background: The induced neural stem cells (iNSCs) held great promises for cell replacement therapy, but iNSCs modulation need improvement. Matrix stiffness could control stem cell fates and might be effective to iNSCs modulations. Here the stiffness of hydrogel matrix on the adhesion, proliferation and differentiation of iNSCs were studied.
Methods: Hyaluronic acid (HA) hydrogels with gradient stiffness were prepared. The structure and stiffness of hydrogels were detected by scanning electron microscopy (SEM) and rheological test. iNSCs were generated from human blood mononuclear cells and cultured in the hydrogels. The cell adhesion, proliferation and differentiation on gradient stiffness hydrogels were examined by CCK-8 test and immunofluorescence staining.
Results: All hydrogels showed typical soft tissue, with the elastic modulus increasing with concentration (0.6-1.8%), ranging from 17 to 250 Pa. The iNSCs maintained growth and differentiation on all gels, but showed different behaviors to different stiffness. On the softer hydrogels, cells grew slowly at first but continuously and fast for long term, tending to differentiate into neurons; while on the harder hydrogels, cells adhered and grew faster at the early stage, tending to differentiate into glia cells after long term culture.
Conclusions: The results suggested that hydrogels stiffness could regulate the key cellular processes of iNSCs. It was important for iNSCs modulation and application in the future.

Keywords:  Hyaluronic acid (HA); differentiation; hydrogel; induced neural stem cells (iNSCs); stiffness

DOI:  https://doi.org/10.21037/atm-21-6189