On-Chip Living-Cell Microarrays for Network Biology

Willaert, Ronnie; Sahli, Hichem

doi:10.5772/20499

Cited by 3 publications

(3 citation statements)

References 134 publications

(120 reference statements)

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“…The inner pH of the cell, as well as aeration, both affect the induction of yeast CYP monooxygenase and CYP reductase [10]. Studies show that a higher dissolved oxygen (DO) level can remarkably improve P450 activity [99,100]. This level was kept at 80% saturation for LCDCA production from C. ropiclais [14].…”

Section: Discussionmentioning

confidence: 99%

Yeast Biotechnology 2.0

Willaert¹

2018

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Research Professor, has an extensive expertise in yeast research (Saccharomyces cerevisiae, S. pastorianus, Candida albicans, and C. glabrata) and single-molecule biophysics (high-resolution microscopy, i.e., confocal laser microscopy, AFM, force spectroscopy, and scanning probe lithography), yeast space biology research and hardware development, protein science (yeast adhesins), cell (yeast) immobilization biotechnology, fermentation technology, and brewing science and technology. vii fermentation

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Section: Discussionmentioning

confidence: 99%

Yeast Biotechnology 2.0

Willaert¹

2018

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“…Additionally, the cell number and density of a given area or volume can be controlled, which allows monitoring of a high spatial and temporal resolution and observation of the dynamic behavior of many cells [93,94]. Parallellisation of experimental conditions and automatization in microfluidic chips has resulted in designs for high-throughput cell screening [95], such as living cell microarrays [96][97][98][99][100][101]. Microfluidic parallelization and single-cell monitoring allows the measurement and averaging of parameters on hundreds of individual cells, compared to measuring parameters of a whole cell population.…”

Section: Microfluidic High-throughput Antifungal Drug Discoverymentioning

confidence: 99%

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

Willaert

2018

Fermentation

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Clinical needs for novel antifungal agents have increased due to the increase of people with a compromised immune system, the appearance of resistant fungi, and infections by unusual yeasts. The search for new molecular targets for antifungals has generated considerable research, especially using modern omics methods (genomics, genome-wide collections of mutants, and proteomics) and bioinformatics approaches. Recently, micro-and nanoscale approaches have been introduced in antifungal drug discovery. Microfluidic platforms have been developed, since they have a number of advantages compared to traditional multiwell-plate screening, such as low reagent consumption, the manipulation of a large number of cells simultaneously and independently, and ease of integrating numerous analytical standard operations and large-scale integration. Automated high-throughput antifungal drug screening is achievable by massive parallel processing. Various microfluidic antimicrobial susceptibility testing (AST) methods have been developed, since they can provide the result in a short time-frame, which is necessary for personalized medicine in the clinic. New nanosensors, based on detecting the nanomotions of cells, have been developed to further decrease the time to test antifungal susceptibility to a few minutes. Finally, nanoparticles (especially, silver nanoparticles) that demonstrated antifungal activity are reviewed.Fermentation 2018, 4, 43 2 of 23 antifungal classes have been reported since 2006 [7]. Current antifungal drugs show some limitations: Amphothericin B (a polyene antibiotic) displays a considerable toxicity and undesirable side effects [8,9], issues with pharmacokinetic properties (such as a short half-life of echinocandins) and activity spectrum, a small number of targets [10,11], and they can interact with other drugs, such as chemotherapy agents and immunosuppressants [12,13]. The last approved antifungal (i.e., anidulafungin) by the European Medicines Agency and the Food and Drug Administration (FDA) dates back to 2006 [14]. There is an urgent need for safer and more effective antifungal drugs. Multiple types of antifungal compounds are in clinical development and these new agents have been recently reviewed [15][16][17].Over the last 20 years, several approaches to antifungal discovery have been explored. The traditional approach seeks, first, to identify active compounds from large compound libraries using a panel of fungal pathogens in standardised assays when possible. In the genetic, genomic, or bioinformatics approach, the objective is, initially, to identify broadly represented targets in fungal pathogens and non-pathogens [18]. In this "target-centric" genomic approach, up-front genetic, bioinformatics, and biochemical target prioritisation is performed and, subsequently, an in vitro-based screening of individual targets is achieved. However, an exceedingly high rate of failure has been observed when applying this approach [19]. Additionally, not all bioactive compounds act through a target-specific mechan...

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“…A cellular microarray consists of a solid support wherein small volumes of different biomolecules and cells can be displayed in defined locations, allowing the multiplexed interrogation of living cells and the analysis of cellular responses [4,5]. Living cell microarrays have been combined with microfluidic bioreactors, which provide multiple advantages for multiplex dynamic analyses and high-throughput screening [4,6]. Cellular arrays are emerging as important tools for functional genomics, drug discovery, toxicology, and stem cell research [4,7,8].…”

Section: Introductionmentioning

confidence: 99%

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

2020

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Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Here, we developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of cell variants (green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones). We combined mechanical patterning in 102 microwells and robotic piezoelectric cell dispensing in the microwells to construct the cell arrays. Robotic yeast cell dispensing of a yeast collection from a multiwell plate to the microfluidic chip microwells was optimized. The developed microfluidic chip and procedure were validated by observing the growth of GFP-tagged yeast clones that are linked to the cell cycle by time-lapse fluorescence microscopy over a few generations. The developed microfluidic technology has the potential to be easily upscaled to a high-density cell array allowing us to perform dynamic proteomics and localizomics experiments.

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On-Chip Living-Cell Microarrays for Network Biology

Cited by 3 publications

References 134 publications

Yeast Biotechnology 2.0

Yeast Biotechnology 2.0

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

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