bims-orenst 2021-07-25 papers

bims-orenst

Biomed News

on Organs-on-chips and engineered stem cell models

Issue of 2021–07–25
seven papers selected by
Joram Mooiweer, University of Groningen

A novel microphysiological colon platform to decipher mechanisms driving human intestinal permeability.
Modeling alcohol-associated liver disease in a human Liver-Chip.
From Model System to Therapy: Scalable Production of Perfusable Vascularized Liver Spheroids in "Open-Top" 384-Well Plate.
Rapid Prototyping of Multilayer Microphysiological Systems.
Human tumor microenvironment chip evaluates the consequences of platelet extravasation and combinatorial antitumor-antiplatelet therapy in ovarian cancer.
Production of Multiple Cell-Laden Microtissue Spheroids with a Biomimetic Hepatic-Lobule-Like Structure.
Usability of Polydimethylsiloxane-Based Microfluidic Devices in Pharmaceutical Research Using Human Hepatocytes.

Cell Mol Gastroenterol Hepatol. 2021 Jul 17. pii: S2352-345X(21)00145-4. [Epub ahead of print]

A novel microphysiological colon platform to decipher mechanisms driving human intestinal permeability.

Athanasia Apostolou, Rohit A Panchakshari, Antara Banerjee, Dimitris V Manatakis, Maria D Paraskevopoulou, Raymond Luc, Galeb Abu-Ali, Alexandra Dimitriou, Carolina Lucchesi, Gauri Kulkarni, Tengku Ibrahim Maulana, Magdalena Kasendra, Jordan S Kerns, Bertram Bleck, Lorna Ewart, Elias S Manolakos, Geraldine A Hamilton, Cosmas Giallourakis, Katia Karalis.

   BACKGROUND & AIMS: The limited availability of organoid systems that mimic the molecular signatures and architecture of human intestinal epithelium has been an impediment to allowing them to be harnessed for the development of therapeutics as well as physiological insights. We developed a microphysiological Organ-on-Chip platform designed to mimic properties of human intestinal epithelium leading to insights into barrier integrity.
METHODS: We combined the human biopsy-derived LGR5+ organoids and Organ-on-Chip technologies to establish a micro-engineered human Colon Intestine-Chip. We characterized the proximity of the model to human tissue and organoids maintained in suspension by RNAseq analysis, and their differentiation to IECs on the Colon Intestine-Chip under variable conditions. Furthermore, organoids from different donors were evaluated to understand variability in the system. Our system was applied to understanding epithelial barrier and characterizing mechanisms driving the cytokine-induced barrier disruption.
RESULTS: Our data highlight the importance of the endothelium and the in vivo tissue-relevant dynamic microenvironment in the Colon Intestine-Chip in the establishment of a tight monolayer of differentiated, polarized organoid-derived intestinal epithelial cells. We confirmed the effect of interferon-gamma (IFNγ) on the colonic barrier and identified reorganization of apical junctional complexes, and induction of apoptosis in the IECs as mediating mechanisms. We demonstrate that in the human Colon Intestine-Chip exposure to interleukin 22 (IL-22) induces disruption of the barrier, unlike its described protective role in experimental colitis in mice.
CONCLUSION: We developed a human Colon Intestine-Chip platform and demonstrated its value in the characterization of the mechanism of action of IL-22 in the human epithelial barrier. This system can be used to elucidate, in a time- and challenge-dependent manner, the mechanism driving the development of leaky gut in humans and to identify associated biomarkers.

Keywords:  Organ-on-Chip; interleukin 22; leaky gut; organoids

DOI:  https://doi.org/10.1016/j.jcmgh.2021.07.004
Cell Rep. 2021 Jul 20. pii: S2211-1247(21)00791-9. [Epub ahead of print]36(3): 109393

Modeling alcohol-associated liver disease in a human Liver-Chip.

Janna C Nawroth, Debora B Petropolis, Dimitris V Manatakis, Tengku Ibrahim Maulana, Gabriel Burchett, Katharina Schlünder, Anke Witt, Abhishek Shukla, Konstantia Kodella, Janey Ronxhi, Gauri Kulkarni, Geraldine Hamilton, Ekihiro Seki, Shelly Lu, Katia C Karalis.

  Alcohol-associated liver disease (ALD) is a global health issue and leads to progressive liver injury, comorbidities, and increased mortality. Human-relevant preclinical models of ALD are urgently needed. Here, we leverage a triculture human Liver-Chip with biomimetic hepatic sinusoids and bile canaliculi to model ALD employing human-relevant blood alcohol concentrations (BACs) and multimodal profiling of clinically relevant endpoints. Our Liver-Chip recapitulates established ALD markers in response to 48 h of exposure to ethanol, including lipid accumulation and oxidative stress, in a concentration-dependent manner and supports the study of secondary insults, such as high blood endotoxin levels. We show that remodeling of the bile canalicular network can provide an in vitro quantitative readout of alcoholic liver toxicity. In summary, we report the development of a human ALD Liver-Chip as a powerful platform for modeling alcohol-induced liver injury with the potential for direct translation to clinical research and evaluation of patient-specific responses.

Keywords:  ALD; ASH; NASH; alcohol; bile canaliculi; digital pathology; fatty liver; liver disease; organ-on-chip; steatosis

DOI:  https://doi.org/10.1016/j.celrep.2021.109393
ACS Biomater Sci Eng. 2021 Jul 12. 7(7): 2964-2972

From Model System to Therapy: Scalable Production of Perfusable Vascularized Liver Spheroids in "Open-Top" 384-Well Plate.

Dawn S Y Lin, Shravanthi Rajasekar, Mandeep Kaur Marway, Boyang Zhang.

  Vasculature is a key component of many biological tissues and helps to regulate a wide range of biological processes. Modeling vascular networks or the vascular interface in organ-on-a-chip systems is an essential aspect of this technology. In many organ-on-a-chip devices, however, the engineered vasculatures are usually designed to be encapsulated inside closed microfluidic channels, making it difficult to physically access or extract the tissues for downstream applications and analysis. One unexploited benefit of tissue extraction is the potential of vascularizing, perfusing, and maturing the tissue in well-controlled, organ-on-a-chip microenvironments and then subsequently extracting that product for in vivo therapeutic implantation. Moreover, for both modeling and therapeutic applications, the scalability of the tissue production process is important. Here we demonstrate the scalable production of perfusable and extractable vascularized tissues in an "open-top" 384-well plate (referred to as IFlowPlate), showing that this system could be used to examine nanoparticle delivery to targeted tissues through the microvascular network and to model vascular angiogenesis. Furthermore, tissue spheroids, such as hepatic spheroids, can be vascularized in a scalable manner and then subsequently extracted for in vivo implantation. This simple multiple-well plate platform could not only improve the experimental throughputs of organ-on-a-chip systems but could potentially help expand the application of model systems to regenerative therapy.

Keywords:  angiogenesis; hydrogel; liver; organ-on-a-chip; tissue spheroids; vasculature

DOI:  https://doi.org/10.1021/acsbiomaterials.0c00236
ACS Biomater Sci Eng. 2021 Jul 12. 7(7): 2949-2963

Rapid Prototyping of Multilayer Microphysiological Systems.

Sanjin Hosic, Adam J Bindas, Marissa L Puzan, Will Lake, Jonathan R Soucy, Fanny Zhou, Ryan A Koppes, David T Breault, Shashi K Murthy, Abigail N Koppes.

  Microfluidic organs-on-chips aim to realize more biorelevant in vitro experiments compared to traditional two-dimensional (2D) static cell culture. Often such devices are fabricated via poly(dimethylsiloxane) (PDMS) soft lithography, which offers benefits (e.g., high feature resolution) along with drawbacks (e.g., prototyping time/costs). Here, we report benchtop fabrication of multilayer, PDMS-free, thermoplastic organs-on-chips via laser cut and assembly with double-sided adhesives that overcome some limitations of traditional PDMS lithography. Cut and assembled chips are economical to prototype ($2 per chip), can be fabricated in parallel within hours, and are Luer compatible. Biocompatibility was demonstrated with epithelial line Caco-2 cells and primary human small intestinal organoids. Comparable to control static Transwell cultures, Caco-2 and organoids cultured on chips formed confluent monolayers expressing tight junctions with low permeability. Caco-2 cells-on-chip differentiated ∼4 times faster, including increased mucus, compared to controls. To demonstrate the robustness of cut and assemble, we fabricated a dual membrane, trilayer chip integrating 2D and 3D compartments with accessible apical and basolateral flow chambers. As proof of concept, we cocultured a human, differentiated monolayer and intact 3D organoids within multilayered contacting compartments. The epithelium exhibited 3D tissue structure and organoids expanded close to the adjacent monolayer, retaining proliferative stem cells over 10 days. Taken together, cut and assemble offers the capability to rapidly and economically manufacture microfluidic devices, thereby presenting a compelling fabrication technique for developing organs-on-chips of various geometries to study multicellular tissues.

Keywords:  adhesive; economical; fabrication; gut chip; intestinal organoid; organ-on-chip; patient-derived cells; primary epithelium; thermoplastic

DOI:  https://doi.org/10.1021/acsbiomaterials.0c00190
Sci Adv. 2021 Jul;pii: eabg5283. [Epub ahead of print]7(30):

Human tumor microenvironment chip evaluates the consequences of platelet extravasation and combinatorial antitumor-antiplatelet therapy in ovarian cancer.

Biswajit Saha, Tanmay Mathur, James J Tronolone, Mithil Chokshi, Giriraj K Lokhande, Amirali Selahi, Akhilesh K Gaharwar, Vahid Afshar-Kharghan, Anil K Sood, Gang Bao, Abhishek Jain.

Platelets extravasate from the circulation into tumor microenvironment, enable metastasis, and confer resistance to chemotherapy in several cancers. Therefore, arresting tumor-platelet cross-talk with effective and atoxic antiplatelet agents in combination with anticancer drugs may serve as an effective cancer treatment strategy. To test this concept, we create an ovarian tumor microenvironment chip (OTME-Chip) that consists of a platelet-perfused tumor microenvironment and which recapitulates platelet extravasation and its consequences. By including gene-edited tumors and RNA sequencing, this organ-on-chip revealed that platelets and tumors interact through glycoprotein VI (GPVI) and tumor galectin-3 under shear. Last, as proof of principle of a clinical trial, we showed that a GPVI inhibitor, Revacept, impairs metastatic potential and improves chemotherapy. Since GPVI is an antithrombotic target that does not impair hemostasis, it represents a safe cancer therapeutic. We propose that OTME-Chip could be deployed to study other vascular and hematological targets in cancer.

DOI: https://doi.org/10.1126/sciadv.abg5283
Adv Mater. 2021 Jul 19. e2102624

Production of Multiple Cell-Laden Microtissue Spheroids with a Biomimetic Hepatic-Lobule-Like Structure.

Gyusik Hong, Jin Kim, Hyeongkwon Oh, Seokhwan Yun, Chul Min Kim, Yun-Mi Jeong, Won-Soo Yun, Jin-Hyung Shim, Ilho Jang, C-Yoon Kim, Songwan Jin.

  The construction of an in vitro 3D cellular model to mimic the human liver is highly desired for drug discovery and clinical applications, such as patient-specific treatment and cell-based therapy in regenerative medicine. However, current bioprinting strategies are limited in their ability to generate multiple cell-laden microtissues with biomimetic structures. This study presents a method for producing hepatic-lobule-like microtissue spheroids using a bioprinting system incorporating a precursor cartridge and microfluidic emulsification system. The multiple cell-laden microtissue spheroids can be successfully generated at a speed of approximately 45 spheroids min-1 and with a uniform diameter. Hepatic and endothelial cells are patterned in a microtissue spheroid with the biomimetic structure of a liver lobule. The spheroids allow long-term culture with high cell viability, and the structural integrity is maintained longer than that of non-structured spheroids. Furthermore, structured spheroids show high MRP2, albumin, and CD31 expression levels. In addition, the in vivo study reveals that structured microtissue spheroids are stably engrafted. These results demonstrate that the method provides a valuable 3D structured microtissue spheroid model with lobule-like constructs and liver functions.

Keywords:  3D bioprinting; hepatic lobules; microtissues; preset extrusions; spheroids; tissue engineering

DOI:  https://doi.org/10.1002/adma.202102624
ACS Biomater Sci Eng. 2021 Jul 20.

Usability of Polydimethylsiloxane-Based Microfluidic Devices in Pharmaceutical Research Using Human Hepatocytes.

Sayaka Deguchi, Masahiro Tsuda, Kaori Kosugi, Ayaka Sakamoto, Natsumi Mimura, Ryosuke Negoro, Emi Sano, Takuro Nobe, Kazuya Maeda, Hiroyuki Kusuhara, Hiroyuki Mizuguchi, Fumiyoshi Yamashita, Yu-Suke Torisawa, Kazuo Takayama.

  A liver-on-a-chip (liver-chip) is a microfluidic device carrying liver cells such as human hepatocytes. It is used to reproduce a part of liver function. Many microfluidic devices are composed of polydimethylsiloxane (PDMS), which is a type of silicone elastomer. PDMS is easy to process and suitable for cell observation, but its high hydrophobicity carries the risk of drug absorption. In this study, we evaluated drug absorption to the PDMS device and investigated the drug responsiveness of human hepatocytes cultured in the PDMS device (hepatocyte-chips). First, the absorption rates of 12 compounds to the PDMS device were measured. The absorption rates of midazolam, bufuralol, cyclosporine A, and verapamil were 92.9, 71.7, 71.4, and 99.6%, respectively, but the other compounds were poorly absorbed. Importantly, the absorption rate of the compounds was correlated with their octanol/water distribution coefficient (log D) values (R2 = 0.76). Next, hepatocyte-chips were used to examine the response to drugs, which are typically used to evaluate hepatic functions. Using the hepatocyte-chips, we could confirm the responsiveness of drugs including cytochrome P450 (CYP) inducers and farnesoid X receptor (FXR) ligands. We believe that our findings will contribute to drug discovery research using PDMS-based liver-chips.

Keywords:  S + log D; drug absorption; human hepatocyte; liver-on-a-chip; organ-on-a-chip; polydimethylsiloxane device

DOI:  https://doi.org/10.1021/acsbiomaterials.1c00642