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Small Methods Nov 2021Few microfluidic devices are used in biomedical labs, despite the obvious potential; reasons given include the devices are rarely made with cell-friendly materials, and... (Review)
Review
Few microfluidic devices are used in biomedical labs, despite the obvious potential; reasons given include the devices are rarely made with cell-friendly materials, and liquids are inaccessibly buried behind solid confining walls. An open microfluidic approach is reviewed in which aqueous circuits with almost any imaginable 2D shape are fabricated in minutes on standard polystyrene Petri dishes by reshaping two liquids (cell-culture media plus an immiscible and bioinert fluorocarbon, FC40). Then, the aqueous phase becomes confined by fluid FC40 walls firmly pinned to the dish by interfacial forces. Such walls can be pierced at any point with pipets and liquids added or removed through them, while flows can be driven actively using external pumps or passively by exploiting local differences in Laplace pressure. As walls are robust, permeable to O plus CO , and transparent, cells are grown in incubators and monitored microscopically as usual. It is hoped that this simple, accessible, and affordable fluid-shaping technology provides bioscientists with an easy entrée into microfluidics.
Topics: Animals; Cell Culture Techniques; Fluorocarbons; Humans; Microfluidic Analytical Techniques
PubMed: 34927960
DOI: 10.1002/smtd.202100724 -
Methods in Molecular Biology (Clifton,... 2016Drosophila melanogaster is an incredibly versatile organism capable of both innate and higher-order behaviors. These behaviors offer not only a way to assay whether or... (Review)
Review
Drosophila melanogaster is an incredibly versatile organism capable of both innate and higher-order behaviors. These behaviors offer not only a way to assay whether or not the animal is physiologically compromised (e.g., feeding, locomotion), but also serve to assess changes in centrally mediated functions. Here we describe several high throughput, reproducible, yet inexpensive and facile behavioral assays for both larval and adult Drosophila. The larval assays all employ an agar substrate in a petri dish; the adult assays are grouped into "vial-based" and "arena-based" paradigms. While these protocols are largely designed to assess individual animals, they are sufficiently rapid that ample numbers can be tested to determine behavioral significance. Importantly, this also allows for one to control for reproductive status, age, and sex, since these factors all have a significant impact on adult behaviors. In general, it is best to designate a dedicated area for any assay, so that lighting conditions are consistent, and all animals should be tested at roughly the same time each day to minimize circadian fluctuations. Temperature and humidity should also be maintained at a constant level to minimize variability in the assays.
Topics: Animals; Benzaldehydes; Biological Assay; Chemotaxis; Circadian Rhythm; Drosophila melanogaster; Embryo, Mammalian; Feeding Behavior; Female; Heptanol; Humidity; Larva; Light; Locomotion; Male; Odorants; Phototaxis; Temperature
PubMed: 27730591
DOI: 10.1007/978-1-4939-6371-3_19 -
Journal of Biomedical Materials... Oct 2021The cell microenvironment such as substrate topology plays an important role in biological processes. In this study, microgrooves were successfully produced on surfaces...
The cell microenvironment such as substrate topology plays an important role in biological processes. In this study, microgrooves were successfully produced on surfaces of both thermoplastic and thermoset polymers using cost-effective techniques for mass production. The micropatterning of thermoplastic polystyrene (PS) petri dish was accomplished efficiently using an in-house developed low-cost hot embossing system. The high replication fidelity of the microgroove with depth and width of 2 μm and spacing of 2 μm was achieved by using silicone rubber as a soft counter mold. This patterned petri dish subsequently served as the cast to replicate the micropattern onto thermoset polydimethylsiloxane (PDMS). It was found that the micropattern increased the hydrophobicity of both PS and PDMS surfaces. The effect of the substrate micropattern on cellular behaviors was preliminarily investigated with untreated and treated PS petri dish as well as PDMS. The results show that the micropattern significantly improved cell adhesion and proliferation for cells cultured on untreated PS petri dish and PDMS substrates. Moreover, the micropattern induced obvious cell alignment along the microgrooves for culturing on all substrates which were studied.
Topics: Cell Adhesion; Cell Culture Techniques; Cell Proliferation; Coated Materials, Biocompatible; Dimethylpolysiloxanes; Hydrophobic and Hydrophilic Interactions; Mesenchymal Stem Cells; Polystyrenes; Surface Properties
PubMed: 33590658
DOI: 10.1002/jbm.b.34811 -
Biomaterials Science Jun 2023Extracellular vesicles (EVs) are lipid-based particles naturally released from cells and recognized as promising drug delivery vehicles for improving therapeutic... (Review)
Review
Extracellular vesicles (EVs) are lipid-based particles naturally released from cells and recognized as promising drug delivery vehicles for improving therapeutic outcomes. Efficient manufacturing of therapeutic EVs have been challenging for their clinical translations. Three-dimensional (3D) cell cultures enabled by biomaterial scaffolds have been used as a platform for improving EV manufacturing compared to conventional methods such as isolation from bodily fluids and standard Petri-dish cell culture. Recent studies on 3D culture derived EV production prove it to enhance the EV yield, functional cargos, and therapeutic efficacies. However, there are still challenges with scaling up 3D cell culture production platforms for industrial use. Hence, there is a high demand for designing, optimizing, and implementing large scale EV manufacturing platforms derived from 3D cell cultures. We will first review the current advances of biomaterial-enabled 3D cell cultures in EV manufacturing, followed by the effect of these 3D cell culture platforms on the EV yield, the EV quality, and therapeutic efficacies. Lastly, we will discuss the key challenges and potential for implementing biomaterial-enabled 3D culture in EV manufacturing for large scale processes in the industrial use.
Topics: Biocompatible Materials; Extracellular Vesicles; Cell Culture Techniques; Cell Culture Techniques, Three Dimensional; Drug Delivery Systems
PubMed: 37222265
DOI: 10.1039/d3bm00469d -
Micromachines Feb 2019In a forest of a hundred thousand trees, no two leaves are alike. Similarly, no two cells in a genetically identical group are the same. This heterogeneity at the... (Review)
Review
In a forest of a hundred thousand trees, no two leaves are alike. Similarly, no two cells in a genetically identical group are the same. This heterogeneity at the single-cell level has been recognized to be vital for the correct interpretation of diagnostic and therapeutic results of diseases, but has been masked for a long time by studying average responses from a population. To comprehensively understand cell heterogeneity, diverse manipulation and comprehensive analysis of cells at the single-cell level are demanded. However, using traditional biological tools, such as petri-dishes and well-plates, is technically challengeable for manipulating and analyzing single-cells with small size and low concentration of target biomolecules. With the development of microfluidics, which is a technology of manipulating and controlling fluids in the range of micro- to pico-liters in networks of channels with dimensions from tens to hundreds of microns, single-cell study has been blooming for almost two decades. Comparing to conventional petri-dish or well-plate experiments, microfluidic single-cell analysis offers advantages of higher throughput, smaller sample volume, automatic sample processing, and lower contamination risk, etc., which made microfluidics an ideal technology for conducting statically meaningful single-cell research. In this review, we will summarize the advances of microfluidics for single-cell manipulation and analysis from the aspects of methods and applications. First, various methods, such as hydrodynamic and electrical approaches, for microfluidic single-cell manipulation will be summarized. Second, single-cell analysis ranging from cellular to genetic level by using microfluidic technology is summarized. Last, we will also discuss the advantages and disadvantages of various microfluidic methods for single-cell manipulation, and then outlook the trend of microfluidic single-cell analysis.
PubMed: 30717128
DOI: 10.3390/mi10020104 -
Parasitology Research Sep 2021Phototaxis is the common behavioral response exhibited by the oncomiracidia of various monogeneans. However, the changes in the oncomiracidial swimming behavior in...
Phototaxis is the common behavioral response exhibited by the oncomiracidia of various monogeneans. However, the changes in the oncomiracidial swimming behavior in response to light cues are not well understood. Here, we investigated the light responses of four monogeneans that are important pathogens in mariculture, namely Benedenia epinepheli, Benedenia seriolae, Neobenedenia girellae, and Heteraxine heterocerca. The swimming trajectory and speed of oncomiracidia of each species were assessed in a glass Petri dish with an LED light placed adjacent to it, based on three different light responses: LED light in the off position (normal swimming), LED light in the on position (phototactic behavior), and immediately (< 5 s) after switching the LED light off (photophobic behavior). The oncomiracidia of all four species exhibited positive phototactic and photophobic responses; however, the change in swimming speed between each response differed among the species. The oncomiracidia of three species (B. epinepheli, N. girellae, and H. heterocerca) exhibited high swimming speed, as a phototactic response; in contrast, the oncomiracidia of B. seriolae exhibited reduced swimming speed when moving toward the light source. Benedenia epinepheli and H. heterocerca exhibited the highest swimming speed during the phototaxis phase, whereas B. seriolae and N. girellae exhibited the highest swimming speed during the photophobic phase. These light responses are considered adaptive traits to increase the chance of encountering and infecting suitable hosts in nature, and such responses could potentially be applied to the control of parasite infections in aquaculture.
Topics: Animals; Aquaculture; Phototaxis; Trematoda
PubMed: 34405279
DOI: 10.1007/s00436-021-07280-y -
Langmuir : the ACS Journal of Surfaces... Nov 2023Hydrophilicity is a requisite attribute for the 2D cell culture substrate's surface, facilitating cell adhesion and spreading. Conventional poly(dimethylsiloxane) (PDMS)...
Hydrophilicity is a requisite attribute for the 2D cell culture substrate's surface, facilitating cell adhesion and spreading. Conventional poly(dimethylsiloxane) (PDMS) microfluidic chips necessitate protein coatings to enhance hydrophilicity; however, this approach is afflicted by issues of transient efficacy, interference with cell analysis, and high costs. This paper presents a protein-free microfluidic chip, termed a "microfluidic Petri dish-chip (MPD-chip)", integrating PDMS as the cover and a tissue culture-treated (TC-treated) Petri dish as the substrate. Microstructures are hot-embossed onto the Petri dish substrate using a silicon mold. This meticulous replication process serves to establish stable flow field dynamics within the chip. A simplified method for irreversible bonding, utilizing plasma activation and silylation, is proposed for affixing the PDMS cover onto the microstructured Petri dish substrate. The prepared composite chip exhibits remarkable tightness, boasting a notable bond strength of 2825 kPa. Furthermore, the composite microfluidic chip demonstrates the capability to withstand flow velocities of at least 200 μL/min, effectively meeting the required injection standards for both cell suspension and culture medium. SH-SY5Y and HeLa cells are cultured dynamically in the MPD-chip and control groups. Outcomes encompassing normalized cell density, cell adhesion area, and cell viability metrics unequivocally highlight the superiority of the MPD-chip in facilitating long-term two-dimensional (2D) cell cultures.
Topics: Humans; Microfluidics; Microfluidic Analytical Techniques; HeLa Cells; Neuroblastoma; Cell Culture Techniques; Proteins
PubMed: 37906157
DOI: 10.1021/acs.langmuir.3c01982 -
Biochip Journal May 2023In vitro model systems have been advanced to recapitulate important physiological features of the target organ in vivo more closely than the conventional cell line... (Review)
Review
In vitro model systems have been advanced to recapitulate important physiological features of the target organ in vivo more closely than the conventional cell line cultures on a petri dish. The advanced organotypic model systems can be used as a complementary or alternative tool for various testing and screening. Numerous data from germ-free animal studies and genome sequencings of clinical samples indicate that human microbiota is an essential part of the human body, but current in vitro model systems rarely include them, which can be one of the reasons for the discrepancy in the tissue phenotypes and outcome of therapeutic intervention between in vivo and in vitro tissues. A coculture model system with appropriate microbes and host cells may have great potential to bridge the gap between the in vitro model and the in vivo counterpart. However, successfully integrating two species in one system introduces new variables to consider and poses new challenges to overcome. This review aims to provide perspectives on the important factors that should be considered for developing organotypic bacterial coculture models. Recent advances in various organotypic bacterial coculture models are highlighted. Finally, challenges and opportunities in developing organotypic microbial coculture models are also discussed.
PubMed: 37363268
DOI: 10.1007/s13206-023-00103-5 -
Biomedicines Apr 2024Stroke is a common neurological disorder, the second leading cause of death, and the third leading cause of disability. Unfortunately, the only approved drug for it is... (Review)
Review
Stroke is a common neurological disorder, the second leading cause of death, and the third leading cause of disability. Unfortunately, the only approved drug for it is tissue plasminogen, but the therapeutic window is limited. In this context, preclinical studies are relevant to better dissect the underlying mechanisms of stroke and for the drug screening of potential therapies. Brain organoids could be relevant in this setting. They are derived from pluripotent stem cells or isolated organ progenitors that differentiate to form an organ-like tissue, exhibiting multiple cell types that self-organize to form a structure not unlike the organ in vivo. Brain organoids mimic many key features of early human brain development at molecular, cellular, structural, and functional levels and have emerged as novel model systems that can be used to investigate human brain diseases including stroke. Brain organoids are a promising and powerful tool for ischemic stroke studies; however, there are a few concerns that need to be addressed, including the lack of vascularization and the many cell types that are typically present in the human brain. The aim of this review is to discuss the potential of brain organoids as a novel model system for studying ischemic stroke, highlighting both the advantages and disadvantages in the use of this technology.
PubMed: 38672231
DOI: 10.3390/biomedicines12040877 -
Science Advances Sep 2020Acoustic tweezers are a promising technology for the biocompatible, precise manipulation of delicate bioparticles ranging from nanometer-sized exosomes to...
Acoustic tweezers are a promising technology for the biocompatible, precise manipulation of delicate bioparticles ranging from nanometer-sized exosomes to millimeter-sized zebrafish larva. However, their widespread usage is hindered by their low compatibility with the workflows in biological laboratories. Here, we present multifunctional acoustic tweezers that can manipulate bioparticles in a disposable Petri dish. Various functionalities including cell patterning, tissue engineering, concentrating particles, translating cells, stimulating cells, and cell lysis are demonstrated. Moreover, leaky surface acoustic wave-based holography is achieved by encoding required phases in electrode profiles of interdigitated transducers. This overcomes the frequency and resolution limits of previous holographic techniques to control three-dimensional acoustic beams in microscale. This study presents a favorable technique for noncontact and label-free manipulation of bioparticles in commonly used Petri dishes. It can be readily adopted by the biological and medical communities for cell studies, tissue generation, and regenerative medicine.
PubMed: 32917678
DOI: 10.1126/sciadv.abb0494