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Micromachines
Special Issues
Microfluidic Platforms for Cell Culture and Investigations
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Microfluidic Platforms for Cell Culture and Investigations

A special issue of Micromachines (ISSN 2072-666X). This special issue belongs to the section "B：Biology and Biomedicine".

Deadline for manuscript submissions: closed (1 May 2020) | Viewed by 28017

Special Issue Editors

Prof. Dr. Nobuyuki Futai

E-Mail Website
Guest Editor
Department of Mechanical Engineering, Shibaura Institute of Technology, 3-7-5 Toyosu, Koto-ku, Tokyo 135-8548, Japan
Interests: mobile cell culture systems; on-chip reconstruction of vascular remodeling; cell-free DNA isolation
Special Issues, Collections and Topics in MDPI journals
Dr. Atsushi Takano

E-Mail Website
Guest Editor
Digital Manufacturing and Design, Singapore University of Technology and Design, 8 Somapah Road, Singapore 487372, Singapore
Interests: mobile cell culture systems; cryopreservation of cells; digital manufacturing of cell-based devices

Special Issue Information

Dear Colleagues,

A cell culture is the result of a collection of techniques for isolating cells from animals or plants and maintaining them in apparatus, instead of using them at their origin. The aim of the cell culture is to control the fluidic, physicochemical, nutritional, scaffold, and other biological requirements for their survival, proliferation, and desired differentiation. Recent advances in microtechnology and tissue engineering have extended the cell culture to a wider range of cell numbers and densities, from single cells to 3D co-cultures, and for a wider range of cell culture cycles, from quick disposable cultures to very-long-term cell cultures. Microfluidic technology has also enabled the integration of various cell culture techniques into a chip, i.e., towards the realization of miniaturized cell-processing systems.

This Special Issue aims to turn the spotlight on microsystems for advanced cell culture. Numerous microfluidic systems for various cell-based assays are already the subject of intensive research, including on-chip cellomics and organ-on-chip, as already covered in other Special Issues in Micromachines. This Special Issue rather focuses on the concepts, devices, and methods for enhanced cell culture, including but not limited to fluidic/temperature/gas level control methods, cell handling such as sorting and preservation, and monitoring of cell culture media, all of which constitute the fundamentals of on-chip cellular assays and processing. In addition, this Special Issue will cover simple and effective microfluidic methods that address physicochemical instability and/or high adoption, and the learning costs that inherently come from microtechnology-origin cell culture systems. Downsizing and improving the traditional cell culture techniques will also open new possibilities in portable and affordable cell processing and cell-based sensing systems that will greatly contribute to precision medicine and accurate environmental monitoring.

Prof. Dr. Nobuyuki Futai
Dr. Atsushi Takano
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Micromachines is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2600 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • Long-Term Culture
  • Hypoxic/Hyperoxic Culture
  • Culture in Gradients
  • Coculture/3D Culture
  • Single-Cell Culture
  • On-Chip Cell Sorting/Harvesting/Passaging
  • On-Chip Culture for Environment Monitoring
  • Cryopreservation

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Published Papers (6 papers)

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Research

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14 pages, 4979 KiB  
Article
Integrated On-Chip 3D Vascular Network Culture under Hypoxia
by Miguel Ángel Olmedo-Suárez, Tomohiro Sekiguchi, Atsushi Takano, Maria del Pilar Cañizares-Macías and Nobuyuki Futai
Micromachines 2020, 11(5), 475; https://doi.org/10.3390/mi11050475 - 30 Apr 2020
Cited by 6 | Viewed by 4407
Abstract
We developed a portable device made of poly(dimethylsiloxane) (PDMS)/polymethylmethacrylate (PMMA) for long-term 3D cell culture of vascular endothelial cells for the development of a vascular network and evaluated the device under different transitions between normoxia and hypoxia with good optical accessibility. The combination [...] Read more.
We developed a portable device made of poly(dimethylsiloxane) (PDMS)/polymethylmethacrylate (PMMA) for long-term 3D cell culture of vascular endothelial cells for the development of a vascular network and evaluated the device under different transitions between normoxia and hypoxia with good optical accessibility. The combination of a nested reservoir device and a bicarbonate/ascorbate buffer system accomplished on-chip incubation with 4.91 ± 0.86% pO2 and 5.19 ± 1.70% pCO2 for up to 10 days. Seventy-two hours of normoxic incubation preceding hypoxic culture increased the cell viability, network formation, and size and stability of the resulting lumens compared with those completely maintained in normoxia for the same total duration. We employed different parameters of the network (e.g., total mesh area, total length, number of branches, among others) for the comparison of different oxygen treatments in the device. The differential effect of hypoxic conditions based on the maturity of the vessels may be used as an external factor to improve vascular development in vitro. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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14 pages, 2658 KiB  
Article
Survival Rate of Cells Sent by a Low Mechanical Load Tube Pump: The “Ring Pump”
by Kaoru Uesugi, Keizo Nishiyama, Koki Hirai, Hiroaki Inoue, Yoichi Sakurai, Yoji Yamada, Takashi Taniguchi and Keisuke Morishima
Micromachines 2020, 11(4), 447; https://doi.org/10.3390/mi11040447 - 23 Apr 2020
Cited by 3 | Viewed by 4049
Abstract
A ring pump (RP) is a useful tool for microchannels and automated cell culturing. We have been developing RPs (a full-press ring pump, FRP; and a mid-press ring pump, MRP). However, damage to cells which were sent by the RP and the MRP [...] Read more.
A ring pump (RP) is a useful tool for microchannels and automated cell culturing. We have been developing RPs (a full-press ring pump, FRP; and a mid-press ring pump, MRP). However, damage to cells which were sent by the RP and the MRP was not investigated, and no other studies have compared the damage to cells between RPs and peristaltic pumps (PPs). Therefore, first, we evaluated the damage to cells that were sent by a small size FRP (s-FRP) and small size MRPs (s-MRPs; gap = 25 or 50 μm, respectively). “Small size” means that the s-FRP and the s-MRPs are suitable for microchannel-scale applications. The survival rate of cells sent by the s-MRPs was higher than those sent by the s-FRP, and less damage caused by the former. Second, we compared the survival rate of cells that were sent by a large size FRP (l-FRP), a large size MRP (l-MRP) (gap = 50 μm) and a PP. “Large size” means that the l-FRP and the l-MRP are suitable for automated cell culture system applications. We could not confirm any differences among the cell survival rates. On the other hand, when cells suspended in Dulbecco’s phosphate-buffered saline solution were circulated with the l-MRP (gap = 50 μm) and the PP, we confirmed a difference in cell survival rate, and less damage caused by the former. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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11 pages, 4637 KiB  
Article
Lab-on-Chip Platform for Culturing and Dynamic Evaluation of Cells Development
by Agnieszka Podwin, Danylo Lizanets, Dawid Przystupski, Wojciech Kubicki, Patrycja Śniadek, Julita Kulbacka, Artur Wymysłowski, Rafał Walczak and Jan A. Dziuban
Micromachines 2020, 11(2), 196; https://doi.org/10.3390/mi11020196 - 14 Feb 2020
Cited by 14 | Viewed by 3123
Abstract
This paper presents a full-featured microfluidic platform ensuring long-term culturing and behavioral analysis of the radically different biological micro-objects. The platform uses all-glass lab-chips and MEMS-based components providing dedicated micro-aquatic habitats for the cells, as well as their intentional disturbances on-chip. Specially developed [...] Read more.
This paper presents a full-featured microfluidic platform ensuring long-term culturing and behavioral analysis of the radically different biological micro-objects. The platform uses all-glass lab-chips and MEMS-based components providing dedicated micro-aquatic habitats for the cells, as well as their intentional disturbances on-chip. Specially developed software was implemented to characterize the micro-objects metrologically in terms of population growth and cells’ size, shape, or migration activity. To date, the platform has been successfully applied for the culturing of freshwater microorganisms, fungi, cancer cells, and animal oocytes, showing their notable population growth, high mobility, and taxis mechanisms. For instance, circa 100% expansion of porcine oocytes cells, as well as nearly five-fold increase in E. gracilis population, has been achieved. These results are a good base to conduct further research on the platform versatile applications. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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11 pages, 3572 KiB  
Article
Unveiling the Potential of Droplet Generation, Sorting, Expansion, and Restoration in Microfluidic Biochips
by Yi-Lung Chiu, Ruchi Ashok Kumar Yadav, Hong-Yuan Huang, Yi-Wen Wang and Da-Jeng Yao
Micromachines 2019, 10(11), 756; https://doi.org/10.3390/mi10110756 - 6 Nov 2019
Cited by 4 | Viewed by 3732
Abstract
Microfluidic biochip techniques are prominently replacing conventional biochemical analyzers by the integration of all functions necessary for biochemical analysis using microfluidics. The microfluidics of droplets offer exquisite control over the size of microliter samples to satisfy the requirements of embryo culture, which might [...] Read more.
Microfluidic biochip techniques are prominently replacing conventional biochemical analyzers by the integration of all functions necessary for biochemical analysis using microfluidics. The microfluidics of droplets offer exquisite control over the size of microliter samples to satisfy the requirements of embryo culture, which might involve a size ranging from picoliter to nanoliter. Polydimethylsiloxane (PDMS) is the mainstream material for the fabrication of microfluidic devices due to its excellent biocompatibility and simplicity of fabrication. Herein, we developed a microfluidic biomedical chip on a PDMS substrate that integrated four key functions—generation of a droplet of an emulsion, sorting, expansion and restoration, which were employed in a mouse embryo system to assess reproductive medicine. The main channel of the designed chip had width of 1200 μm and height of 500 μm. The designed microfluidic chips possessed six sections—cleaved into three inlets and three outlets—to study the key functions with five-day embryo culture. The control part of the experiment was conducted with polystyrene (PS) beads (100 μm), the same size as the murine embryos, for the purpose of testing. The outcomes of our work illustrate that the rate of success of the static droplet culture group (87.5%) is only slightly less than that of a conventional group (95%). It clearly demonstrates that a droplet-based microfluidic system can produce a droplet in a volume range from picoliter to nanoliter. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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10 pages, 1983 KiB  
Article
High-Throughput Platform for Efficient Chemical Transfection, Virus Packaging, and Transduction
by Jianxiong Zhang, Yawei Hu, Xiaoqing Wang, Peng Liu and Xiaofang Chen
Micromachines 2019, 10(6), 387; https://doi.org/10.3390/mi10060387 - 10 Jun 2019
Cited by 5 | Viewed by 3871
Abstract
Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, [...] Read more.
Intracellular gene delivery is normally required to study gene functions. A versatile platform able to perform both chemical transfection and viral transduction to achieve efficient gene modification in most cell types is needed. Here we demonstrated that high throughput chemical transfection, virus packaging, and transduction can be conducted efficiently on our previously developed superhydrophobic microwell array chip (SMAR-chip). A total of 169 chemical transfections were successfully performed on the chip in physically separated microwells through a few simple steps, contributing to the convenience of DNA delivery and media change on the SMAR-chip. Efficiencies comparable to the traditional transfection in multi-well plates (~65%) were achieved while the manual operations were largely reduced. Two transfection procedures, the dry method amenable for the long term storage of the transfection material and the wet method for higher efficiencies were developed. Multiple transfections in a scheduled manner were performed to further increase the transfection efficiencies or deliver multiple genes at different time points. In addition, high throughput virus packaging integrated with target cell transduction were also proved which resulted in a transgene expression efficiency of >70% in NIH 3T3 cells. In summary, the SMAR-chip based high throughput gene delivery is efficient and versatile, which can be used for large scale genetic modifications in a variety of cell types. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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Review

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20 pages, 3521 KiB  
Review
In Vitro Blood–Brain Barrier-Integrated Neurological Disorder Models Using a Microfluidic Device
by Jin-Ha Choi, Mallesh Santhosh and Jeong-Woo Choi
Micromachines 2020, 11(1), 21; https://doi.org/10.3390/mi11010021 - 24 Dec 2019
Cited by 19 | Viewed by 8165
Abstract
The blood–brain barrier (BBB) plays critical role in the human physiological system such as protection of the central nervous system (CNS) from external materials in the blood vessel, including toxicants and drugs for several neurological disorders, a critical type of human disease. Therefore, [...] Read more.
The blood–brain barrier (BBB) plays critical role in the human physiological system such as protection of the central nervous system (CNS) from external materials in the blood vessel, including toxicants and drugs for several neurological disorders, a critical type of human disease. Therefore, suitable in vitro BBB models with fluidic flow to mimic the shear stress and supply of nutrients have been developed. Neurological disorder has also been investigated for developing realistic models that allow advance fundamental and translational research and effective therapeutic strategy design. Here, we discuss introduction of the blood–brain barrier in neurological disorder models by leveraging a recently developed microfluidic system and human organ-on-a-chip system. Such models could provide an effective drug screening platform and facilitate personalized therapy of several neurological diseases. Full article
(This article belongs to the Special Issue Microfluidic Platforms for Cell Culture and Investigations)
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