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From the Lab-on-a-Chip to the Organ-on-a-Chip

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A special issue of Genes (ISSN 2073-4425). This special issue belongs to the section "Technologies and Resources for Genetics".

Deadline for manuscript submissions: closed (30 April 2018) | Viewed by 110620

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Special Issue Editors

Prof. Dr. Rimantas Kodzius

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Guest Editor

iSmart, Materials Genome Institute, Shanghai University (SHU), Shanghai 200000, China

Dr. Frank Schulze

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Guest Editor

Federal Institute for Risk Assessment (BfR), 10115 Berlin, Germany

Prof. Dr. Marlon R. Schneider

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Co-Guest Editor

Federal Institute for Risk Assessment (BfR), 10115 Berlin, Germany

Dr. Xinghua Gao

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Co-Guest Editor

iSmart, Materials Genome Institute, Shanghai University (SHU), Shanghai 200000, China

Special Issue Information

Dear Colleagues,

Human health is of great importance to all of us; yet, a great extent of the present understanding of human physiology, that is, the function of organs, tissues, and cells in an organism, is based on the use of cell lines in a 2D monolayer culture. There are countless cell lines available from various origins, such as human blood, different organs, and even pathological tissues, such as tumors.

Although of immense value in past research, conventional 2D cell cultures are strongly limited in recreating the complex interactions of different cell types, tissues and organs within higher organisms (e.g., vertebrates). The communication of cells through direct cell–cell contact, and by the release of chemokines, are also skewed in 2D cell culture since 3D structures are missing and the ratio of cells to the volume of surrounding fluid (media) is non-physiological. Furthermore, tissue-specific parameters, such as pH, pO₂, extracellular matrix composition, or the presence of mechanical forces, are underrepresented in monolayer cell cultures.

Thus, the translation of the data gained in conventional cell cultures into complex organisms is hampered, a disadvantage that is apparent in many fields, such as regenerative medicine or toxicology. To compensate for these shortcomings, animal experiments are widely used, especially in risk assessment or the testing of chemicals and novel drugs. Here, animal testing provides some information on drug activity; however, in any case, drugs are then tested in subsequent phases (clinical trials) on human probands.

On the other hand, the culture of cells outside the human body is challenging and requires specific knowledge regarding the accompanying limitations. While it is the standard to grow cells in 2D monolayer cultures, which only poorly resemble physiological situations, there have been many attempts to cultivate them in 3D to improve their biological relevance. The further transition from 3D cell culture models to organ-on-a-chip (OoC) systems allows for the recreation of a physiological environment that resembles the parameters of the tissues of interest and also allows the co-culture of different tissue-specific cell types in 2D or 3D.

Recent advances in Microelectromechanical Systems (MEMS) enable the construction of OoCs with structures in the range of nano- and micrometers. Various materials can be utilized—from silica to ceramics, glass, metals or different polymers (some with refractory properties similar to glass). Since cells require a water-based environment, microfluidics are especially suitable for their cultivation. Microfluidics offers precise control and manipulation of fluids with many advantages over classic bioreactor systems. It is possible to design growth chambers and flow channels in practically any size and shape, while the volumes of reagents, samples and cell organoids are comparably small. The risk of contamination can be reduced, for example, by utilizing disposable chips. Due to a change in surface-to-volume ratios, the speed of chemical reactions is increased when compared to the macro-scale, resulting in enhanced heat transfer and lower energy requirements.

Advances in miniaturization and manufacturing processes have led to the availability of a plethora of cheap sensors that allow for easy control, thus leading to the rapid automatization of OoCs.

We are facing the successful marriage between cell biology and microfluidic chips, leading to the infancy of OoC. We are sure that the importance of OoC technology will lead to the successful development of various OoCs and their interconnection in one platform, also termed body-on-a-chip.

We anticipate that the further development and standardization of OoCs will result in faster and cheaper drug development, without the need of using laboratory animals for testing. Further OoC use will facilitate a more fundamental understanding of cell function and cell communication. OoC technology may be suitable, not only for the study of human cells, but also for other animal cells, prokaryotes, or even viruses.

Let us share the latest developments of OoC with a series of original high-quality articles and reviews in this Special Issue of Genes.

Prof. Dr. Rimantas Kodzius
Dr. Ing. Frank Schulze
Prof. Dr. Marlon R. Schneider
Dr. Xinghua Gao
Guest Editors

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Keywords

Disease models
Regulatory toxicology
Risk assessment
Drug development
Toxin testing
In vitro testing
Organ-on-a-chip (OoC)
Cell Culture/ Cell Co-Culture
3D cell-culture
Cell interaction
Cell microenvironment
Micro cell culture analog (uCCA)
Biomechanics
Microelectrochemical Systems (MEMS)
Miniaturization
Microfluidics
Microsensors
Material science

Published Papers (13 papers)

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Editorial

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148 KiB

Open AccessEditorial

Organ-on-Chip Technology: Current State and Future Developments

by Rimantas Kodzius, Frank Schulze, Xinghua Gao and Marlon R. Schneider

Genes 2017, 8(10), 266; https://doi.org/10.3390/genes8100266 - 11 Oct 2017

Cited by 28 | Viewed by 5489

Abstract

In the early days of pharmacy, the development of new drugs was frequently achieved by restless chemists who worked solitarily, day by day for years [...]
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(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

Research

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16 pages, 1845 KiB

Open AccessArticle

High-Throughput Incubation and Quantification of Agglutination Assays in a Microfluidic System

by David Castro, David Conchouso, Rimantas Kodzius, Arpys Arevalo and Ian G. Foulds

Genes 2018, 9(6), 281; https://doi.org/10.3390/genes9060281 - 04 Jun 2018

Cited by 11 | Viewed by 3703

Abstract

In this paper, we present a two-phase microfluidic system capable of incubating and quantifying microbead-based agglutination assays. The microfluidic system is based on a simple fabrication solution, which requires only laboratory tubing filled with carrier oil, driven by negative pressure using a syringe pump. We provide a user-friendly interface, in which a pipette is used to insert single droplets of a 1.25-µL volume into a system that is continuously running and therefore works entirely on demand without the need for stopping, resetting or washing the system. These assays are incubated by highly efficient passive mixing with a sample-to-answer time of 2.5 min, a 5–10-fold improvement over traditional agglutination assays. We study system parameters such as channel length, incubation time and flow speed to select optimal assay conditions, using the streptavidin-biotin interaction as a model analyte quantified using optical image processing. We then investigate the effect of changing the concentration of both analyte and microbead concentrations, with a minimum detection limit of 100 ng/mL. The system can be both low- and high-throughput, depending on the rate at which assays are inserted. In our experiments, we were able to easily produce throughputs of 360 assays per hour by simple manual pipetting, which could be increased even further by automation and parallelization. Agglutination assays are a versatile tool, capable of detecting an ever-growing catalog of infectious diseases, proteins and metabolites. A system such as this one is a step towards being able to produce high-throughput microfluidic diagnostic solutions with widespread adoption. The development of analytical techniques in the microfluidic format, such as the one presented in this work, is an important step in being able to continuously monitor the performance and microfluidic outputs of organ-on-chip devices. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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15 pages, 9820 KiB

Open AccessArticle

Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications

by Tobias Grix, Alicia Ruppelt, Alexander Thomas, Anna-Klara Amler, Benjamin P. Noichl, Roland Lauster and Lutz Kloke

Genes 2018, 9(4), 176; https://doi.org/10.3390/genes9040176 - 22 Mar 2018

Cited by 106 | Viewed by 9249

Abstract

Many tissue models have been developed to mimic liver-specific functions for metabolic and toxin conversion in in vitro assays. Most models represent a 2D environment rather than a complex 3D structure similar to native tissue. To overcome this issue, spheroid cultures have become the gold standard in tissue engineering. Unfortunately, spheroids are limited in size due to diffusion barriers in their dense structures, limiting nutrient and oxygen supply. Recent developments in bioprinting techniques have enabled us to engineer complex 3D structures with perfusion-enabled channel systems to ensure nutritional supply within larger, densely-populated tissue models. In this study, we present a proof-of-concept for the feasibility of bioprinting a liver organoid by combining HepaRG and human stellate cells in a stereolithographic printing approach, and show basic characterization under static cultivation conditions. Using standard tissue engineering analytics, such as immunohistology and qPCR, we found higher albumin and cytochrome P₄₅₀ 3A4 (CYP3A4) expression in bioprinted liver tissues compared to monolayer controls over a two-week cultivation period. In addition, the expression of tight junctions, liver-specific bile transporter multidrug resistance-associated protein 2 (MRP2), and overall metabolism (glucose, lactate, lactate dehydrogenase (LDH)) were found to be stable. Furthermore, we provide evidence for the perfusability of the organoids’ intrinsic channel system. These results motivate new approaches and further development in liver tissue engineering for advanced organ-on-a-chip applications and pharmaceutical developments. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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10 pages, 2042 KiB

Open AccessArticle

Skin-on-a-Chip: Transepithelial Electrical Resistance and Extracellular Acidification Measurements through an Automated Air-Liquid Interface

by Frank A. Alexander, Jr., Sebastian Eggert and Joachim Wiest

Genes 2018, 9(2), 114; https://doi.org/10.3390/genes9020114 - 21 Feb 2018

Cited by 55 | Viewed by 9739

Abstract

Skin is a critical organ that plays a crucial role in defending the internal organs of the body. For this reason, extensive work has gone into creating artificial models of the epidermis for in vitro skin toxicity tests. These tissue models, called reconstructed human epidermis (RhE), are used by researchers in the pharmaceutical, cosmetic, and environmental arenas to evaluate skin toxicity upon exposure to xenobiotics. Here, we present a label-free solution that leverages the use of the intelligent mobile lab for in vitro diagnostics (IMOLA-IVD), a noninvasive, sensor-based platform, to monitor the transepithelial electrical resistance (TEER) of RhE models and adherent cells cultured on porous membrane inserts. Murine fibroblasts cultured on polycarbonate membranes were first used as a test model to optimize procedures using a custom BioChip encapsulation design, as well as dual fluidic configurations, for continuous and automated perfusion of membrane-bound cultures. Extracellular acidification rate (EAR) and TEER of membrane-bound L929 cells were monitored. The developed protocol was then used to monitor the TEER of MatTek EpiDerm^TM RhE models over a period of 48 h. TEER and EAR measurements demonstrated that the designed system is capable of maintaining stable cultures on the chip, monitoring metabolic parameters, and revealing tissue breakdown over time. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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11 pages, 2792 KiB

Open AccessArticle

Embedded Disposable Functionalized Electrochemical Biosensor with a 3D-Printed Flow Cell for Detection of Hepatic Oval Cells (HOCs)

by Samar Damiati, Martin Peacock, Stefan Leonhardt, Laila Damiati, Mohammed A. Baghdadi, Holger Becker, Rimantas Kodzius and Bernhard Schuster

Genes 2018, 9(2), 89; https://doi.org/10.3390/genes9020089 - 14 Feb 2018

Cited by 31 | Viewed by 5853

Abstract

Hepatic oval cells (HOCs) are considered the progeny of the intrahepatic stem cells that are found in a small population in the liver after hepatocyte proliferation is inhibited. Due to their small number, isolation and capture of these cells constitute a challenging task for immunosensor technology. This work describes the development of a 3D-printed continuous flow system and exploits disposable screen-printed electrodes for the rapid detection of HOCs that over-express the OV6 marker on their membrane. Multiwall carbon nanotube (MWCNT) electrodes have a chitosan film that serves as a scaffold for the immobilization of oval cell marker antibodies (anti-OV6-Ab), which enhance the sensitivity of the biomarker and makes the designed sensor specific for oval cells. The developed sensor can be easily embedded into the 3D-printed flow cell to allow cells to be exposed continuously to the functionalized surface. The continuous flow is intended to increase capture of most of the target cells in the specimen. Contact angle measurements were performed to characterize the nature and quality of the modified sensor surface, and electrochemical measurements (cyclic voltammetry (CV) and square wave voltammetry (SWV)) were performed to confirm the efficiency and selectivity of the fabricated sensor to detect HOCs. The proposed method is valuable for capturing rare cells and could provide an effective tool for cancer diagnosis and detection. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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4011 KiB

Open AccessArticle

High-Throughput Study of the Effects of Celastrol on Activated Fibroblast-Like Synoviocytes from Patients with Rheumatoid Arthritis

by Zhengyu Fang, Dongyi He, Bo Yu, Feng Liu, Jianping Zuo, Yuxia Li, Qi Lin, Xiaodong Zhou and Qingwen Wang

Genes 2017, 8(9), 221; https://doi.org/10.3390/genes8090221 - 06 Sep 2017

Cited by 28 | Viewed by 5407

Abstract

Celastrol, a natural triterpene, exhibits potential anti-inflammatory activity in a variety of inflammatory diseases. The present study aimed to investigate its biological effect on activated fibroblast-like synoviocytes (FLSs) from patients with rheumatoid arthritis (RA). The primary FLSs of the synovial tissues were obtained from synovial biopsies of patients with RA. The normal human FLS line (HFLS) was used as a control. After the RA–FLSs and HFLSs were treated with or without celastrol, various approaches, including the WST-1 assay, transwell assay, real-time PCR and ELISA analysis, were performed to estimate proliferation, invasion and expression of pro-inflammatory cytokines of the RA–FLSs. Microarray analysis was performed to screen for differentially expressed genes in RA–FLSs before and after celastrol treatment. The results showed that treatment of celastrol attenuated both the proliferation and invasion of the RA–FLSs. The expression of several chemokine genes, including CCL2, CXCL10, CXCL12, CCR2 and CXCR4, was significantly changed after celastrol treatment. The genes involved in the NF-κB signaling pathway appeared to be regulated by celastrol. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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5920 KiB

Open AccessArticle

3D Microstructure Inhibits Mesenchymal Stem Cells Homing to the Site of Liver Cancer Cells on a Microchip

by Xingyuan Yang, Xinyue Xu, Yuan Zhang, Weijia Wen and Xinghua Gao

Genes 2017, 8(9), 218; https://doi.org/10.3390/genes8090218 - 01 Sep 2017

Cited by 10 | Viewed by 4858

Abstract

The cell microenvironment consists of multiple types of biophysical and biochemical factors, and represents a complex integrated system that is variable in both time and space. Studies show that changes in biochemical and biophysical factors in cell microenvironments result in significant changes in cellular forms and functions, especially for stem cells. Mesenchymal stem cells (MSCs) are derived from adult stem cells of the mesoderm and play an important role in tissue engineering, regenerative medicine and even cancer therapy. Furthermore, it is found that MSCs can interact with multiple types of tumor cells. The interaction is reflected as two totally different aspects. The negative aspect is that MSCs manifest as tumor-associated fibroblasts and could induce migration of cancer cells and promote tumor formation. On the other hand, MSCs can home to sites of the tumor microenvironment, directionally migrate toward tumor cells and cause tumor cell apoptosis. In this study, we designed and made a simple microfluidic chip for cell co-culture, and studied stem cell homing behavior in the interaction between MSCs and liver cancer cells. Moreover, by etching a three-dimensional microstructure on the base and adding transforming growth factor-β (TGF-β) in the co-culture environment, we studied the impact of biophysical and biochemical factors on stem cell homing behavior, and the causes of such impact. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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Review

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29 pages, 3041 KiB

Open AccessReview

Development of Microplatforms to Mimic the In Vivo Architecture of CNS and PNS Physiology and Their Diseases

by John Saliba, Arij Daou, Samar Damiati, Jessica Saliba, Marwan El-Sabban and Rami Mhanna

Genes 2018, 9(6), 285; https://doi.org/10.3390/genes9060285 - 06 Jun 2018

Cited by 24 | Viewed by 5958

Abstract

Understanding the mechanisms that govern nervous tissues function remains a challenge. In vitro two-dimensional (2D) cell culture systems provide a simplistic platform to evaluate systematic investigations but often result in unreliable responses that cannot be translated to pathophysiological settings. Recently, microplatforms have emerged to provide a better approximation of the in vivo scenario with better control over the microenvironment, stimuli and structure. Advances in biomaterials enable the construction of three-dimensional (3D) scaffolds, which combined with microfabrication, allow enhanced biomimicry through precise control of the architecture, cell positioning, fluid flows and electrochemical stimuli. This manuscript reviews, compares and contrasts advances in nervous tissues-on-a-chip models and their applications in neural physiology and disease. Microplatforms used for neuro-glia interactions, neuromuscular junctions (NMJs), blood-brain barrier (BBB) and studies on brain cancer, metastasis and neurodegenerative diseases are addressed. Finally, we highlight challenges that can be addressed with interdisciplinary efforts to achieve a higher degree of biomimicry. Nervous tissue microplatforms provide a powerful tool that is destined to provide a better understanding of neural health and disease. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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28 pages, 3064 KiB

Open AccessReview

Synthesis of Biomaterials Utilizing Microfluidic Technology

by Xiaohong Wang, Jinfeng Liu, Peizhou Wang, Andrew DeMello, Lingyan Feng, Xiaoli Zhu, Weijia Wen, Rimantas Kodzius and Xiuqing Gong

Genes 2018, 9(6), 283; https://doi.org/10.3390/genes9060283 - 05 Jun 2018

Cited by 44 | Viewed by 6222

Abstract

Recently, microfluidic technologies have attracted an enormous amount of interest as potential new tools for a large range of applications including materials synthesis, chemical and biological detection, drug delivery and screening, point-of-care diagnostics, and in-the-field analysis. Their ability to handle extremely small volumes of fluids is accompanied by additional benefits, most notably, rapid and efficient mass and heat transfer. In addition, reactions performed within microfluidic systems are highly controlled, meaning that many advanced materials, with uniform and bespoke properties, can be synthesized in a direct and rapid manner. In this review, we discuss the utility of microfluidic systems in the synthesis of materials for a variety of biological applications. Such materials include microparticles or microcapsules for drug delivery, nanoscale materials for medicine or cellular assays, and micro- or nanofibers for tissue engineering. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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36 pages, 3631 KiB

Open AccessReview

Journey into Bone Models: A Review

by Julia Scheinpflug, Moritz Pfeiffenberger, Alexandra Damerau, Franziska Schwarz, Martin Textor, Annemarie Lang and Frank Schulze

Genes 2018, 9(5), 247; https://doi.org/10.3390/genes9050247 - 10 May 2018

Cited by 77 | Viewed by 11122

Abstract

Bone is a complex tissue with a variety of functions, such as providing mechanical stability for locomotion, protection of the inner organs, mineral homeostasis and haematopoiesis. To fulfil these diverse roles in the human body, bone consists of a multitude of different cells and an extracellular matrix that is mechanically stable, yet flexible at the same time. Unlike most tissues, bone is under constant renewal facilitated by a coordinated interaction of bone-forming and bone-resorbing cells. It is thus challenging to recreate bone in its complexity in vitro and most current models rather focus on certain aspects of bone biology that are of relevance for the research question addressed. In addition, animal models are still regarded as the gold-standard in the context of bone biology and pathology, especially for the development of novel treatment strategies. However, species-specific differences impede the translation of findings from animal models to humans. The current review summarizes and discusses the latest developments in bone tissue engineering and organoid culture including suitable cell sources, extracellular matrices and microfluidic bioreactor systems. With available technology in mind, a best possible bone model will be hypothesized. Furthermore, the future need and application of such a complex model will be discussed. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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17 pages, 1118 KiB

Open AccessReview

Cell-Free Approaches in Synthetic Biology Utilizing Microfluidics

by Samar Damiati, Rami Mhanna, Rimantas Kodzius and Eva-Kathrin Ehmoser

Genes 2018, 9(3), 144; https://doi.org/10.3390/genes9030144 - 06 Mar 2018

Cited by 44 | Viewed by 9970

Abstract

Synthetic biology is a rapidly growing multidisciplinary branch of science which aims to mimic complex biological systems by creating similar forms. Constructing an artificial system requires optimization at the gene and protein levels to allow the formation of entire biological pathways. Advances in cell-free synthetic biology have helped in discovering new genes, proteins, and pathways bypassing the complexity of the complex pathway interactions in living cells. Furthermore, this method is cost- and time-effective with access to the cellular protein factory without the membrane boundaries. The freedom of design, full automation, and mimicking of in vivo systems reveal advantages of synthetic biology that can improve the molecular understanding of processes, relevant for life science applications. In parallel, in vitro approaches have enhanced our understanding of the living system. This review highlights the recent evolution of cell-free gene design, proteins, and cells integrated with microfluidic platforms as a promising technology, which has allowed for the transformation of the concept of bioprocesses. Although several challenges remain, the manipulation of biological synthetic machinery in microfluidic devices as suitable ‘homes’ for in vitro protein synthesis has been proposed as a pioneering approach for the development of new platforms, relevant in biomedical and diagnostic contexts towards even the sensing and monitoring of environmental issues. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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24 pages, 2566 KiB

Open AccessReview

Microfluidic Devices for Drug Delivery Systems and Drug Screening

by Samar Damiati, Uday B. Kompella, Safa A. Damiati and Rimantas Kodzius

Genes 2018, 9(2), 103; https://doi.org/10.3390/genes9020103 - 16 Feb 2018

Cited by 255 | Viewed by 20283

Abstract

Microfluidic devices present unique advantages for the development of efficient drug carrier particles, cell-free protein synthesis systems, and rapid techniques for direct drug screening. Compared to bulk methods, by efficiently controlling the geometries of the fabricated chip and the flow rates of multiphase fluids, microfluidic technology enables the generation of highly stable, uniform, monodispersed particles with higher encapsulation efficiency. Since the existing preclinical models are inefficient drug screens for predicting clinical outcomes, microfluidic platforms might offer a more rapid and cost-effective alternative. Compared to 2D cell culture systems and in vivo animal models, microfluidic 3D platforms mimic the in vivo cell systems in a simple, inexpensive manner, which allows high throughput and multiplexed drug screening at the cell, organ, and whole-body levels. In this review, the generation of appropriate drug or gene carriers including different particle types using different configurations of microfluidic devices is highlighted. Additionally, this paper discusses the emergence of fabricated microfluidic cell-free protein synthesis systems for potential use at point of care as well as cell-, organ-, and human-on-a-chip models as smart, sensitive, and reproducible platforms, allowing the investigation of the effects of drugs under conditions imitating the biological system. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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2026 KiB

Open AccessReview

Air Quality Effects on Human Health and Approaches for Its Assessment through Microfluidic Chips

by Frank Schulze, Xinghua Gao, Darius Virzonis, Samar Damiati, Marlon R. Schneider and Rimantas Kodzius

Genes 2017, 8(10), 244; https://doi.org/10.3390/genes8100244 - 27 Sep 2017

Cited by 76 | Viewed by 10613

Abstract

Air quality depends on the various gases and particles present in it. Both natural phenomena and human activities affect the cleanliness of air. In the last decade, many countries experienced an unprecedented industrial growth, resulting in changing air quality values, and correspondingly, affecting our life quality. Air quality can be accessed by employing microchips that qualitatively and quantitatively determine the present gases and dust particles. The so-called particular matter 2.5 (PM2.5) values are of high importance, as such small particles can penetrate the human lung barrier and enter the blood system. There are cancer cases related to many air pollutants, and especially to PM2.5, contributing to exploding costs within the healthcare system. We focus on various current and potential future air pollutants, and propose solutions on how to protect our health against such dangerous substances. Recent developments in the Organ-on-Chip (OoC) technology can be used to study air pollution as well. OoC allows determination of pollutant toxicity and speeds up the development of novel pharmaceutical drugs. Full article

(This article belongs to the Special Issue From the Lab-on-a-Chip to the Organ-on-a-Chip)

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