bims-orenst 2022-03-20 papers

bims-orenst

Biomed News

on Organs-on-chips and engineered stem cell models

Issue of 2022–03–20
seven papers selected by

A microphysiological model of human trophoblast invasion during implantation.
Ectopic Lymphoid Follicle Formation and Human Seasonal Influenza Vaccination Responses Recapitulated in an Organ-on-a-Chip.
Real-time monitoring of immediate drug response and adaptation upon repeated treatment in a microfluidic chip system.
Development of Microfluidic, Serum-Free Bronchial Epithelial Cells-on-a-Chip to Facilitate a More Realistic In vitro Testing of Nanoplastics.
A Multi-Organ-on-Chip Approach to Investigate How Oral Exposure to Metals Can Cause Systemic Toxicity Leading to Langerhans Cell Activation in Skin.
Vascularized lung cancer model for evaluating the promoted transport of anticancer drugs and immune cells in an engineered tumor microenvironment.
Biomimetic hydrogel supports initiation and growth of patient-derived breast tumor organoids.

Nat Commun. 2022 Mar 15. 13(1): 1252

A microphysiological model of human trophoblast invasion during implantation.

Ju Young Park, Sneha Mani, Geremy Clair, Heather M Olson, Vanessa L Paurus, Charles K Ansong, Cassidy Blundell, Rachel Young, Jessica Kanter, Scott Gordon, Alex Y Yi, Monica Mainigi, Dan Dongeun Huh.

Successful establishment of pregnancy requires adhesion of an embryo to the endometrium and subsequent invasion into the maternal tissue. Abnormalities in this critical process of implantation and placentation lead to many pregnancy complications. Here we present a microenigneered system to model a complex sequence of orchestrated multicellular events that plays an essential role in early pregnancy. Our implantation-on-a-chip is capable of reconstructing the three-dimensional structural organization of the maternal-fetal interface to model the invasion of specialized fetal extravillous trophoblasts into the maternal uterus. Using primary human cells isolated from clinical specimens, we demonstrate in vivo-like directional migration of extravillous trophoblasts towards a microengineered maternal vessel and their interactions with the endothelium necessary for vascular remodeling. Through parametric variation of the cellular microenvironment and proteomic analysis of microengineered tissues, we show the important role of decidualized stromal cells as a regulator of extravillous trophoblast migration. Furthermore, our study reveals previously unknown effects of pre-implantation maternal immune cells on extravillous trophoblast invasion. This work represents a significant advance in our ability to model early human pregnancy, and may enable the development of advanced in vitro platforms for basic and clinical research of human reproduction.

DOI: https://doi.org/10.1038/s41467-022-28663-4
Adv Sci (Weinh). 2022 Mar 14. e2103241

Ectopic Lymphoid Follicle Formation and Human Seasonal Influenza Vaccination Responses Recapitulated in an Organ-on-a-Chip.

Girija Goyal, Pranav Prabhala, Gautam Mahajan, Bruce Bausk, Tal Gilboa, Liangxia Xie, Yunhao Zhai, Roey Lazarovits, Adam Mansour, Min Sun Kim, Aditya Patil, Danielle Curran, Jaclyn M Long, Sanjay Sharma, Abidemi Junaid, Limor Cohen, Thomas C Ferrante, Oren Levy, Rachelle Prantil-Baun, David R Walt, Donald E Ingber.

  Lymphoid follicles (LFs) are responsible for generation of adaptive immune responses in secondary lymphoid organs and form ectopically during chronic inflammation. A human model of ectopic LF formation will provide a tool to understand LF development and an alternative to non-human primates for preclinical evaluation of vaccines. Here, it is shown that primary human blood B- and T-lymphocytes autonomously assemble into ectopic LFs when cultured in a 3D extracellular matrix gel within one channel of a two-channel organ-on-a-chip microfluidic device. Superfusion via a parallel channel separated by a microporous membrane is required for LF formation and prevents lymphocyte autoactivation. These germinal center-like LFs contain B cells expressing Activation-Induced Cytidine Deaminase and exhibit plasma cell differentiation upon activation. To explore their utility for seasonal vaccine testing, autologous monocyte-derived dendritic cells are integrated into LF Chips. The human LF chips demonstrate improved antibody responses to split virion influenza vaccination compared to 2D cultures, which are enhanced by a squalene-in-water emulsion adjuvant, and this is accompanied by increases in LF size and number. When inoculated with commercial influenza vaccine, plasma cell formation and production of anti-hemagglutinin IgG are observed, as well as secretion of cytokines similar to vaccinated humans over clinically relevant timescales.

Keywords:  antibody; germinal center; lymph node; organ chip; vaccine

DOI:  https://doi.org/10.1002/advs.202103241
Arch Toxicol. 2022 Mar 19.

Real-time monitoring of immediate drug response and adaptation upon repeated treatment in a microfluidic chip system.

Anastasiia Zuieva, Suzan Can, Franziska Boelke, Stefanie Reuter, Sebastian Schattscheider, Elfi Töpfer, Anika Westphal, Ralf Mrowka, Stefan Wölfl.

  Microfluidic tissue culture and organ-on-a-chip models provide efficient tools for drug testing in vivo and are considered to become the basis of in vitro test systems to analyze drug response, drug interactions and toxicity to complement and reduce animal testing. A major limitation is the efficient recording of drug action. Here we present an efficient experimental setup that allows long-term cultivation of cells in a microfluidic system in combination with continuous recording of luciferase reporter gene expression. The system combines a sensitive cooled luminescence camera system in combination with a custom build miniaturized incubation chamber. The setup allows to monitor time-dependent activation, but also the end of drug response. Repeated activation and recovery as well as varying durations of drug treatment periods can be monitored, and different modes of drug activity can be visualized.

Keywords:  Luciferase; Microfluidic culture; Organ-on-a-chip system; Real-time monitoring

DOI:  https://doi.org/10.1007/s00204-022-03272-8
Front Toxicol. 2021 ;3 735331

Development of Microfluidic, Serum-Free Bronchial Epithelial Cells-on-a-Chip to Facilitate a More Realistic In vitro Testing of Nanoplastics.

Govind Gupta, Srikanth Vallabani, Romain Bordes, Kunal Bhattacharya, Bengt Fadeel.

  Most cell culture models are static, but the cellular microenvironment in the body is dynamic. Here, we established a microfluidic-based in vitro model of human bronchial epithelial cells in which cells are stationary, but nutrient supply is dynamic, and we used this system to evaluate cellular uptake of nanoparticles. The cells were maintained in fetal calf serum-free and bovine pituitary extract-free cell culture medium. BEAS-2B, an immortalized, non-tumorigenic human cell line, was used as a model and the cells were grown in a chip within a microfluidic device and were briefly infused with amorphous silica (SiO2) nanoparticles or polystyrene (PS) nanoparticles of similar primary sizes but with different densities. For comparison, tests were also performed using static, multi-well cultures. Cellular uptake of the fluorescently labeled particles was investigated by flow cytometry and confocal microscopy. Exposure under dynamic culture conditions resulted in higher cellular uptake of the PS nanoparticles when compared to static conditions, while uptake of SiO2 nanoparticles was similar in both settings. The present study has shown that it is feasible to grow human lung cells under completely animal-free conditions using a microfluidic-based device, and we have also found that cellular uptake of PS nanoparticles aka nanoplastics is highly dependent on culture conditions. Hence, traditional cell cultures may not accurately reflect the uptake of low-density particles, potentially leading to an underestimation of their cellular impact.

Keywords:  alternative methods; in vitro; microfluidics; nanoplastics; nanotoxicology

DOI:  https://doi.org/10.3389/ftox.2021.735331
Front Toxicol. 2021 ;3 824825

A Multi-Organ-on-Chip Approach to Investigate How Oral Exposure to Metals Can Cause Systemic Toxicity Leading to Langerhans Cell Activation in Skin.

Jasper J Koning, Charlotte T Rodrigues Neves, Katharina Schimek, Maria Thon, Sander W Spiekstra, Taco Waaijman, Tanja D de Gruijl, Susan Gibbs.

  Investigating systemic toxicity in vitro is still a huge challenge. Here, a multi-organ-on-chip approach is presented as a typical case of topical exposure of oral mucosa to metals, which are known to activate the immune system and in turn may result in skin inflammation. Reconstructed human gingiva (RHG) and reconstructed human skin containing MUTZ-3-derived Langerhans cells (MUTZ-LC) in the epidermis (RHS-LC) were incorporated into a HUMIMIC Chip3plus, connected by dynamic flow and cultured for a total period of 72 h. Three independent experiments were performed each with an intra-experiment replicate in order to assess the donor and technical variations. After an initial culture period of 24 h to achieve stable dynamic culture conditions, nickel sulfate was applied topically to RHG for 24 h, and LC activation (maturation and migration) was determined in RHS-LC after an additional 24 h incubation time. A stable dynamic culture of RHG and RHS-LC was achieved as indicated by the assessment of glucose uptake, lactate production, and lactate dehydrogenase release into the microfluidics compartment. Nickel exposure resulted in no major histological changes within RHG or RHS-LC, or cytokine release into the microfluidics compartment, but did result in an increased activation of LC as observed by the increased mRNA levels of CD1a, CD207, HLA-DR, and CD86 in the dermal compartment (hydrogel of RHS-LC (PCR)). This is the first study to describe systemic toxicity and immune cell activation in a multi-organ setting and can provide a framework for studying other organoids in the future.

Keywords:  Langerhans cell; nickel; organ on a chip; organotypic; reconstructed human gingiva; reconstructed human skin

DOI:  https://doi.org/10.3389/ftox.2021.824825
Adv Healthc Mater. 2022 Mar 14. e2102581

Vascularized lung cancer model for evaluating the promoted transport of anticancer drugs and immune cells in an engineered tumor microenvironment.

Dasom Kim, Kyeong Seob Hwang, Eun U Seo, Suyeong Seo, Byung Chul Lee, Nakwon Choi, Jonghoon Choi, Hong Nam Kim.

  The tumor microenvironment (TME) is the environment around the tumor, including blood vessels, immune cells, fibroblasts, signaling molecules, and the extracellular matrix (ECM). Owing to its component interactions, the TME influences tumor growth and drug delivery in a highly complex manner. Although several vascularized cancer models have been developed to mimic the TME in vitro, these models cannot comprehensively reflect blood vessel-tumor spheroid interactions. Here, we present a method for inducing controlled tumor angiogenesis by engineering the microenvironment. The interstitial flow direction regulates the direction of capillary sprouting, showing that angiogenesis occurs in the opposite direction of flow, while the existence of lung fibroblasts affects the continuity and lumen formation of sprouted capillaries. The vascularized tumor model shows enhanced delivery of anticancer drugs and immune cells to the tumor spheroids because of the perfusable vascular networks. The possibility of capillary embolism using anticancer drug-conjugated liquid metal nanoparticles was investigated using the vascularized tumor model. This vascularized tumor platform can aid in the development of effective anticancer drugs and cancer immunotherapy. This article is protected by copyright. All rights reserved.

Keywords:  angiogenesis; drug delivery; immune cell transport; tumor spheroid; vascularization

DOI:  https://doi.org/10.1002/adhm.202102581
Nat Commun. 2022 Mar 18. 13(1): 1466

Biomimetic hydrogel supports initiation and growth of patient-derived breast tumor organoids.

Elisabeth Prince, Jennifer Cruickshank, Wail Ba-Alawi, Kelsey Hodgson, Jillian Haight, Chantal Tobin, Andrew Wakeman, Alona Avoulov, Valentina Topolskaia, Mitchell J Elliott, Alison P McGuigan, Hal K Berman, Benjamin Haibe-Kains, David W Cescon, Eugenia Kumacheva.

Patient-derived tumor organoids (PDOs) are a highly promising preclinical model that recapitulates the histology, gene expression, and drug response of the donor patient tumor. Currently, PDO culture relies on basement-membrane extract (BME), which suffers from batch-to-batch variability, the presence of xenogeneic compounds and residual growth factors, and poor control of mechanical properties. Additionally, for the development of new organoid lines from patient-derived xenografts, contamination of murine host cells poses a problem. We propose a nanofibrillar hydrogel (EKGel) for the initiation and growth of breast cancer PDOs. PDOs grown in EKGel have histopathologic features, gene expression, and drug response that are similar to those of their parental tumors and PDOs in BME. In addition, EKGel offers reduced batch-to-batch variability, a range of mechanical properties, and suppressed contamination from murine cells. These results show that EKGel is an improved alternative to BME matrices for the initiation, growth, and maintenance of breast cancer PDOs.

DOI: https://doi.org/10.1038/s41467-022-28788-6