In comparison to traditional in vitro cell culture in Petri dishes or well plates, cell culture in microfluidic-based devices enables better control over chemical and physical environments, higher levels of experimental automation, and a reduction in experimental materials Over the past decade, the advantages associated with cell culturing in microfluidic-based platforms have garnered significant interest and have led to a plethora of studies for high throughput cell assays, organs-on-a-chip applications, temporal signaling studies, and cell sorting A clear concern for performing cell culture in microfluidic-based devices is deciding on a technique to deliver and pump media to cells that are encased in a microfluidic device In this review, we summarize recent advances in pumping techniques for microfluidic cell culture and discuss their advantages and possible drawbacks The ultimate goal of our review is to distill the large body of information available related to pumps for microfluidic cell culture in an effort to assist current and potential users of microfluidic-based devices for advanced in vitro cellular studies

Pumps for microfluidic cell culture.

Monika Fuxreiter,† Agnes Toth-Petroczy,‡ Daniel A. Kraut, Andreas T. Matouschek, Roderick Y. H. Lim, Bin Xue, Lukasz Kurgan, and Vladimir N. Uversky* †MTA-DE Momentum Laboratory of Protein Dynamics, Department of Biochemistry and Molecular Biology, University of Debrecen, Nagyerdei krt. 98, H-4032 Debrecen, Hungary ‡Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 7610001, Israel Department of Chemistry, Villanova University, 800 East Lancaster Avenue, Villanova, Pennsylvania 19085, United States Section of Molecular Genetics and Microbiology, Institute for Cellular & Molecular Biology, The University of Texas at Austin, 2506 Speedway, Austin, Texas 78712, United States Biozentrum and the Swiss Nanoscience Institute, University of Basel, Klingelbergstrasse 70, CH-4056 Basel, Switzerland Department of Cell Biology, Microbiology and Molecular Biology, College of Fine Arts and Sciences, and Department of Molecular Medicine and USF Health Byrd Alzheimer’s Research Institute, Morsani College of Medicine, University of South Florida, Tampa, Florida 33612, United States Department of Electrical and Computer Engineering, University of Alberta, Edmonton, AB T6G 2R3, Canada Institute for Biological Instrumentation, Russian Academy of Sciences, 142290 Pushchino, Moscow Region 119991, Russia

https://pubs.acs.org/doi/pdfplus/10.1021/cr4007329

Disordered Proteinaceous Machines

Cationic polymers are commonly used to transfect mammalian cells, but their mechanisms of DNA delivery are unknown. This study seeks to decipher the mechanism by which plasmid DNA delivered by a class of cationic polymers traffics to and enters the nucleus. While studies have been performed to elucidate the mechanism of naked plasmid DNA (pDNA) import into the nuclei of mammalian cells, our objectives were to determine the effects of polymer complexation on pDNA nuclear import and the impact of polymer structure on that import. We have performed studies in whole cells and in isolated nuclei using flow cytometry and confocal microscopy to characterize how polymer–DNA complexes (polyplexes) are able to deliver their pDNA cargo to the nuclei of their target cells. The polymers tested herein include (i) linear poly(ethylenimine) (JetPEI), a polyamine, and (ii) two poly(glycoamidoamine)s (PGAAs), polyamines that contain carbohydrate moieties (meso-galactarate, Glycofect (G4), and l-tartarate, T4) within their ...

Exploring the mechanism of plasmid DNA nuclear internalization with polymer-based vehicles.

In a diffusion-based molecular communication network, transmitters and receivers communicate by using signalling molecules (or ligands) in a fluid medium. This paper assumes that the transmitter uses different chemical reactions to generate different emission patterns of signalling molecules to represent different transmission symbols, and the receiver consists of receptors. When the signalling molecules arrive at the receiver, they may react with the receptors to form ligand-receptor complexes. Our goal is to study the demodulation in this setup assuming that the transmitter and receiver are synchronised. We derive an optimal demodulator using the continuous history of the number of complexes at the receiver as the input to the demodulator. We do that by first deriving a communication model which includes the chemical reactions in the transmitter, diffusion in the transmission medium and the ligand-receptor process in the receiver. This model, which takes the form of a continuous-time Markov process, captures the noise in the receiver signal due to the stochastic nature of chemical reactions and diffusion. We then adopt a maximum a posteriori framework and use Bayesian filtering to derive the optimal demodulator. We use numerical examples to illustrate the properties of this optimal demodulator.

A Markovian Approach to the Optimal Demodulation of Diffusion-Based Molecular Communication Networks

The ability to control vascular endothelial growth factor (VEGF) signaling offers promising therapeutic potential for vascular diseases and cancer. Despite this promise, VEGF-targeted therapies are not clinically effective for many pathologies, such as breast cancer. VEGFR1 has recently emerged as a predictive biomarker for anti-VEGF efficacy, implying a functional VEGFR1 role beyond its classically defined decoy receptor status. Here we introduce a computational approach that accurately predicts cellular responses elicited via VEGFR1 signaling. Aligned with our model prediction, we show empirically that VEGFR1 promotes macrophage migration through PLCγ and PI3K pathways and promotes macrophage proliferation through a PLCγ pathway. These results provide new insight into the basic function of VEGFR1 signaling while offering a computational platform to quantify signaling of any receptor.

/pdf/vegfr1-promotes-cell-migration-and-proliferation-through-173wnydjj8.pdf

VEGFR1 promotes cell migration and proliferation through PLCγ and PI3K pathways

This paper introduces the concept of phase-locking analysis of oscillatory cellular signaling systems to elucidate biochemical circuit architecture. Phase-locking is a physical phenomenon that refers to a response mode in which system output is synchronized to a periodic stimulus; in some instances, the number of responses can be fewer than the number of inputs, indicative of skipped beats. While the observation of phase-locking alone is largely independent of detailed mechanism, we find that the properties of phase-locking are useful for discriminating circuit architectures because they reflect not only the activation but also the recovery characteristics of biochemical circuits. Here, this principle is demonstrated for analysis of a G-protein coupled receptor system, the M3 muscarinic receptor-calcium signaling pathway, using microfluidic-mediated periodic chemical stimulation of the M3 receptor with carbachol and real-time imaging of resulting calcium transients. Using this approach we uncovered the potential importance of basal IP3 production, a finding that has important implications on calcium response fidelity to periodic stimulation. Based upon our analysis, we also negated the notion that the Gq-PLC interaction is switch-like, which has a strong influence upon how extracellular signals are filtered and interpreted downstream. Phase-locking analysis is a new and useful tool for model revision and mechanism elucidation; the method complements conventional genetic and chemical tools for analysis of cellular signaling circuitry and should be broadly applicable to other oscillatory pathways.

/pdf/phase-locked-signals-elucidate-circuit-architecture-of-an-4gs08ic5xa.pdf

Phase-locked signals elucidate circuit architecture of an oscillatory pathway

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Phase-locked signals elucidate circuit architecture of an oscillatory pathway