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12 pages, 4871 KiB

Open AccessArticle

Integrated Microfluidic–Electromagnetic System to Probe Single-Cell Magnetotaxis in Microconfinement

by Brianna Bradley, Juan Gomez-Cruz and Carlos Escobedo

Bioengineering 2023, 10(9), 1034; https://doi.org/10.3390/bioengineering10091034 - 01 Sep 2023

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Magnetotactic bacteria have great potential for use in biomedical and environmental applications due to the ability to direct their navigation with a magnetic field. Applying and accurately controlling a magnetic field within a microscopic region during bacterial magnetotaxis studies at the single-cell level [...] Read more.

Magnetotactic bacteria have great potential for use in biomedical and environmental applications due to the ability to direct their navigation with a magnetic field. Applying and accurately controlling a magnetic field within a microscopic region during bacterial magnetotaxis studies at the single-cell level is challenging due to bulky microscope components and the inherent curvilinear field lines produced by commonly used bar magnets. In this paper, a system that integrates microfluidics and electromagnetic coils is presented for generating a linear magnetic field within a microenvironment compatible with microfluidics, enabling magnetotaxis analysis of groups or single microorganisms on-chip. The platform, designed and optimised via finite element analysis, is integrated into an inverted fluorescent microscope, enabling visualisation of bacteria at the single-cell level in microfluidic devices. The electromagnetic coils produce a linear magnetic field throughout a central volume where the microfluidic device containing the magnetotactic bacteria is located. The magnetic field, at this central position, can be accurately controlled from 1 to 10 mT, which is suitable for directing the navigation of magnetotactic bacteria. Potential heating of the microfluidic device from the operating coils was evaluated up to 2.5 A, corresponding to a magnetic field of 7.8 mT, for 10 min. The maximum measured heating was 8.4 °C, which enables analysis without altering the magnetotaxis behaviour or the average swimming speed of the bacteria. Altogether, this work provides a design, characterisation and experimental test of an integrated platform that enables the study of individual bacteria confined in microfluidics, under linear and predictable magnetic fields that can be easily and accurately applied and controlled. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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14 pages, 2966 KiB

Open AccessArticle

Fiber-Based SERS-Fluidic Polymeric Platforms for Improved Optical Analysis of Liquids

by Caterina Credi, Caterina Dallari, Sara Nocentini, Gabriele Gatta, Elena Bianchi, Diederik S. Wiersma and Francesco S. Pavone

Bioengineering 2023, 10(6), 676; https://doi.org/10.3390/bioengineering10060676 - 01 Jun 2023

Viewed by 1202

Abstract

Downsizing surface-enhanced Raman spectroscopy (SERS) within microfluidic devices has opened interesting perspectives for the development of low-cost and portable (bio)sensors for the optical analysis of liquid samples. Despite the research efforts, SERS-fluidic devices still rely either on the use of expensive bulky set-ups [...] Read more.

Downsizing surface-enhanced Raman spectroscopy (SERS) within microfluidic devices has opened interesting perspectives for the development of low-cost and portable (bio)sensors for the optical analysis of liquid samples. Despite the research efforts, SERS-fluidic devices still rely either on the use of expensive bulky set-ups or on polymeric devices giving spurious background signals fabricated via expensive manufacturing processes. Here, polymeric platforms integrating fluidics and optics were fabricated with versatile designs allowing easy coupling with fiber-based Raman systems. For the first time, anti-fouling photocurable perfluoropolyether (PFPE) was explored for high-throughput SERS-integrating chip fabrication via replica molding of negative stamps obtained through standard and advanced fabrication processes. The PFPE devices comprised networks of channels for fluid handling and for optical fiber housing with multiple orientations. Embedded microfeatures were used to control the relative positioning of the fibers, thus guaranteeing the highest signal delivering and collection. The feasibility of PFPE devices as fiber-based SERS fluidic platforms was demonstrated through the straightforward acquisition of Raman-SERS spectra of a mixture of gold nanoparticles as SERS substrates with rhodamine 6G (Rh6G) at decreasing concentrations. In the presence of high-performing gold nanostars, the Rh6G signal was detectable at dilutions down to the nanomolar level even without tight focusing and working at low laser power—a key aspect for analyte detection in real-world biomedical and environmental applications. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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13 pages, 1534 KiB

Open AccessArticle

Establishment of a Perfusion Process with Antibody-Producing CHO Cells Using a 3D-Printed Microfluidic Spiral Separator with Web-Based Flow Control

by Jana Schellenberg, Michaela Dehne, Ferdinand Lange, Thomas Scheper, Dörte Solle and Janina Bahnemann

Bioengineering 2023, 10(6), 656; https://doi.org/10.3390/bioengineering10060656 - 28 May 2023

Cited by 2 | Viewed by 1989

Abstract

Monoclonal antibodies are increasingly dominating the market for human therapeutic and diagnostic agents. For this reason, continuous methods—such as perfusion processes—are being explored and optimized in an ongoing effort to increase product yields. Unfortunately, many established cell retention devices—such as tangential flow filtration—rely [...] Read more.

Monoclonal antibodies are increasingly dominating the market for human therapeutic and diagnostic agents. For this reason, continuous methods—such as perfusion processes—are being explored and optimized in an ongoing effort to increase product yields. Unfortunately, many established cell retention devices—such as tangential flow filtration—rely on membranes that are prone to clogging, fouling, and undesirable product retention at high cell densities. To circumvent these problems, in this work, we have developed a 3D-printed microfluidic spiral separator for cell retention, which can readily be adapted and replaced according to process conditions (i.e., a plug-and-play system) due to the fast and flexible 3D printing technique. In addition, this system was also expanded to include automatic flushing, web-based control, and notification via a cellphone application. This set-up constitutes a proof of concept that was successful at inducing a stable process operation at a viable cell concentration of 10–17 × 10⁶ cells/mL in a hybrid mode (with alternating cell retention and cell bleed phases) while significantly reducing both shear stress and channel blockage. In addition to increasing efficiency to nearly 100%, this microfluidic device also improved production conditions by successfully separating dead cells and cell debris and increasing cell viability within the bioreactor. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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17 pages, 3846 KiB

Open AccessArticle

A Human Ovarian Tumor & Liver Organ-on-Chip for Simultaneous and More Predictive Toxo-Efficacy Assays

by Arianna Fedi, Chiara Vitale, Marco Fato and Silvia Scaglione

Bioengineering 2023, 10(2), 270; https://doi.org/10.3390/bioengineering10020270 - 18 Feb 2023

Cited by 4 | Viewed by 2667

Abstract

In oncology, the poor success rate of clinical trials is becoming increasingly evident due to the weak predictability of preclinical assays, which either do not recapitulate the complexity of human tissues (i.e., in vitro tests) or reveal species-specific outcomes (i.e., animal testing). Therefore, [...] Read more.

In oncology, the poor success rate of clinical trials is becoming increasingly evident due to the weak predictability of preclinical assays, which either do not recapitulate the complexity of human tissues (i.e., in vitro tests) or reveal species-specific outcomes (i.e., animal testing). Therefore, the development of novel approaches is fundamental for better evaluating novel anti-cancer treatments. Here, a multicompartmental organ-on-chip (OOC) platform was adopted to fluidically connect 3D ovarian cancer tissues to hepatic cellular models and resemble the systemic cisplatin administration for contemporarily investigating drug efficacy and hepatotoxic effects in a physiological context. Computational fluid dynamics was performed to impose capillary-like blood flows and predict cisplatin diffusion. After a cisplatin concentration screening using 2D/3D tissue models, cytotoxicity assays were conducted in the multicompartmental OOC and compared with static co-cultures and dynamic single-organ models. A linear decay of SKOV-3 ovarian cancer and HepG2 liver cell viability was observed with increasing cisplatin concentration. Furthermore, 3D ovarian cancer models showed higher drug resistance than the 2D model in static conditions. Most importantly, when compared to clinical therapy, the experimental approach combining 3D culture, fluid-dynamic conditions, and multi-organ connection displayed the most predictive toxicity and efficacy results, demonstrating that OOC-based approaches are reliable 3Rs alternatives in preclinic. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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18 pages, 2537 KiB

Open AccessArticle

Cultivation and Imaging of S. latissima Embryo Monolayered Cell Sheets Inside Microfluidic Devices

by Thomas Clerc, Samuel Boscq, Rafaele Attia, Gabriele S. Kaminski Schierle, Bénédicte Charrier and Nino F. Läubli

Bioengineering 2022, 9(11), 718; https://doi.org/10.3390/bioengineering9110718 - 21 Nov 2022

Cited by 1 | Viewed by 2048

Abstract

The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth [...] Read more.

The culturing and investigation of individual marine specimens in lab environments is crucial to further our understanding of this highly complex ecosystem. However, the obtained results and their relevance are often limited by a lack of suitable experimental setups enabling controlled specimen growth in a natural environment while allowing for precise monitoring and in-depth observations. In this work, we explore the viability of a microfluidic device for the investigation of the growth of the alga Saccharina latissima to enable high-resolution imaging by confining the samples, which usually grow in 3D, to a single 2D plane. We evaluate the specimen’s health based on various factors such as its growth rate, cell shape, and major developmental steps with regard to the device’s operating parameters and flow conditions before demonstrating its compatibility with state-of-the-art microscopy imaging technologies such as the skeletonisation of the specimen through calcofluor white-based vital staining of its cell contours as well as the immunolocalisation of the specimen’s cell wall. Furthermore, by making use of the on-chip characterisation capabilities, we investigate the influence of altered environmental illuminations on the embryonic development using blue and red light. Finally, live tracking of fluorescent microspheres deposited on the surface of the embryo permits the quantitative characterisation of growth at various locations of the organism. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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14 pages, 3568 KiB

Open AccessArticle

Droplet Microfluidics Enables Tracing of Target Cells at the Single-Cell Transcriptome Resolution

by Yang Liu, Shiyu Wang, Menghua Lyu, Run Xie, Weijin Guo, Ying He, Xuyang Shi, Yang Wang, Jingyu Qi, Qianqian Zhu, Hui Zhang, Tao Luo, Huaying Chen, Yonggang Zhu, Xuan Dong, Zida Li, Ying Gu, Longqi Liu, Xun Xu and Ya Liu

Bioengineering 2022, 9(11), 674; https://doi.org/10.3390/bioengineering9110674 - 10 Nov 2022

Cited by 5 | Viewed by 2312

Abstract

The rapid promotion of single-cell omics in various fields has begun to help solve many problems encountered in research, including precision medicine, prenatal diagnosis, and embryo development. Meanwhile, single-cell techniques are also constantly updated with increasing demand. For some specific target cells, the [...] Read more.

The rapid promotion of single-cell omics in various fields has begun to help solve many problems encountered in research, including precision medicine, prenatal diagnosis, and embryo development. Meanwhile, single-cell techniques are also constantly updated with increasing demand. For some specific target cells, the workflow from droplet screening to single-cell sequencing is a preferred option and should reduce the impact of operation steps, such as demulsification and cell recovery. We developed an all-in-droplet method integrating cell encapsulation, target sorting, droplet picoinjection, and single-cell transcriptome profiling on chips to achieve labor-saving monitoring of TCR-T cells. As a proof of concept, in this research, TCR-T cells were encapsulated, sorted, and performed single-cell transcriptome sequencing (scRNA-seq) by injecting reagents into droplets. It avoided the tedious operation of droplet breakage and re-encapsulation between droplet sorting and scRNA-seq. Moreover, convenient device operation will accelerate the progress of chip marketization. The strategy achieved an excellent recovery performance of single-cell transcriptome with a median gene number over 4000 and a cross-contamination rate of 8.2 ± 2%. Furthermore, this strategy allows us to develop a device with high integrability to monitor infused TCR-T cells, which will promote the development of adoptive T cell immunotherapy and their clinical application. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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17 pages, 3557 KiB

Open AccessArticle

Development of Liver-on-Chip Integrating a Hydroscaffold Mimicking the Liver’s Extracellular Matrix

by Taha Messelmani, Anne Le Goff, Zied Souguir, Victoria Maes, Méryl Roudaut, Elodie Vandenhaute, Nathalie Maubon, Cécile Legallais, Eric Leclerc and Rachid Jellali

Bioengineering 2022, 9(9), 443; https://doi.org/10.3390/bioengineering9090443 - 05 Sep 2022

Cited by 10 | Viewed by 3191

Abstract

The 3Rs guidelines recommend replacing animal testing with alternative models. One of the solutions proposed is organ-on-chip technology in which liver-on-chip is one of the most promising alternatives for drug screening and toxicological assays. The main challenge is to achieve the relevant in [...] Read more.

The 3Rs guidelines recommend replacing animal testing with alternative models. One of the solutions proposed is organ-on-chip technology in which liver-on-chip is one of the most promising alternatives for drug screening and toxicological assays. The main challenge is to achieve the relevant in vivo-like functionalities of the liver tissue in an optimized cellular microenvironment. Here, we investigated the development of hepatic cells under dynamic conditions inside a 3D hydroscaffold embedded in a microfluidic device. The hydroscaffold is made of hyaluronic acid and composed of liver extracellular matrix components (galactosamine, collagen I/IV) with RGDS (Arg-Gly-Asp-Ser) sites for cell adhesion. The HepG2/C3A cell line was cultured under a flow rate of 10 µL/min for 21 days. After seeding, the cells formed aggregates and proliferated, forming 3D spheroids. The cell viability, functionality, and spheroid integrity were investigated and compared to static cultures. The results showed a 3D aggregate organization of the cells up to large spheroid formations, high viability and albumin production, and an enhancement of HepG2 cell functionalities. Overall, these results highlighted the role of the liver-on-chip model coupled with a hydroscaffold in the enhancement of cell functions and its potential for engineering a relevant liver model for drug screening and disease study. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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12 pages, 632 KiB

Open AccessPerspective

Boosting the Clinical Translation of Organ-on-a-Chip Technology

by David Caballero, Rui L. Reis and Subhas C. Kundu

Bioengineering 2022, 9(10), 549; https://doi.org/10.3390/bioengineering9100549 - 14 Oct 2022

Cited by 4 | Viewed by 1884

Abstract

Organ-on-a-chip devices have become a viable option for investigating critical physiological events and responses; this technology has matured substantially, and many systems have been reported for disease modeling or drug screening over the last decade. Despite the wide acceptance in the academic community, [...] Read more.

Organ-on-a-chip devices have become a viable option for investigating critical physiological events and responses; this technology has matured substantially, and many systems have been reported for disease modeling or drug screening over the last decade. Despite the wide acceptance in the academic community, their adoption by clinical end-users is still a non-accomplished promise. The reasons behind this difficulty can be very diverse but most likely are related to the lack of predictive power, physiological relevance, and reliability necessary for being utilized in the clinical area. In this Perspective, we briefly discuss the main attributes of organ-on-a-chip platforms in academia and how these characteristics impede their easy translation to the clinic. We also discuss how academia, in conjunction with the industry, can contribute to boosting their adoption by proposing novel design concepts, fabrication methods, processes, and manufacturing materials, improving their standardization and versatility, and simplifying their manipulation and reusability. Full article

(This article belongs to the Special Issue Microfluidics and Miniaturized Systems in Bioengineering)

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