The influence of entropic crowding in cell monolayers

Ippolito, Alberto; Deshpande, V.S.

doi:10.1016/j.bpj.2022.08.030

Cited by 5 publications

(3 citation statements)

References 42 publications

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“…This state can also be inferred from the presence of actin bundles visible along the cell periphery (Figure c, bottom; center); such bundles became exclusively highlighted in the glioma-CMV group. The increase in glioma membrane interactions promotes spatial confinement, with the glioma cells adopting an elongated configuration (Figure a) that resembles the nematic ordering of liquid crystals. , Similar to the phase transitions in liquid crystals subject to shear force, elongated cells in motion experience a torque that reassembles their structural proteins, reshapes their cytoskeleton and alters their polarization state. , …”

Section: Resultsmentioning

confidence: 99%

Tumor-Derived Membrane Vesicles Restrain Migration in Gliomas By Altering Collective Polarization

Jhunjhunwala,

Yu,

Kuo

et al. 2023

ACS Appl. Bio Mater.

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Mechanobiology is a cornerstone in physiology. However, its role in biomedical applications remains considerably undermined. In this study, we employed cell membrane vesicles (CMVs), which are currently being used as nanodrug carriers, as tactile cues for mechano-regulation of collective cell behaviors. Gliomas, which are among the most resilient brain tumors and have a low patient survival rate, were used as the cell model. We observed that mechanical responses due to the application of glioma-or microglia-derived CMVs resulted in the doubling of the traction stress of glioma cell collectives with a 10-fold increase in the CMV concentration. Glioma-CMVs constrained cell protrusions and hindered their collective migration, with the migration speed of such cells declining by almost 40% compared to the untreated cells. We speculated that the alteration of collective polarization leads to migration speed changes, and this phenomenon was elucidated using the cellular Potts model. In addition to intracellular force modulation and cytoskeletal reorganization, glioma-CMVs altered drug diffusion within glioma spheroids by downregulating the mechano-signaling protein YAP-1 while also marginally enhancing the associated apoptotic events. Our results suggest that glioma-CMVs can be applied as an adjuvant to current treatment approaches to restrict tumor invasion and enhance the penetration of reagents within tumors. Considering the broad impact of mechano-transduction on cell functions, the regulation of cell mechanics through CMVs can provide a foundation for alternative therapeutic strategies.

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Section: Resultsmentioning

confidence: 99%

Tumor-Derived Membrane Vesicles Restrain Migration in Gliomas By Altering Collective Polarization

Jhunjhunwala,

Yu,

Kuo

et al. 2023

ACS Appl. Bio Mater.

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“…Moreover, the LID domain of AK3L1 predominates in its effect on enzyme activity, as it undergoes a significant movement during the catalytic cycle. This study is an important step toward a better understanding of how the congested milieu of the physiological interior might regulate the gamut of processes that may seemingly be perturbed by the local crowding environment. − …”

Section: Discussionmentioning

confidence: 99%

Deeper Insights into Mixed Crowding through Enzyme Activity, Dynamics, and Crowder Diffusion

Singh,

Gupta,

Rastogi

et al. 2024

J. Phys. Chem. B

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Given the fact that the cellular interior is crowded by many different kinds of macromolecules, it is important that in vitro studies be carried out in the presence of mixed crowder systems. In this regard, we have used binary crowders formed by the combination of some of the commonly used crowding agents, namely, Ficoll 70, Dextran 70, Dextran 40, and PEG 8000 (PEG 8), to study how these affect enzyme activity, dynamics, and crowder diffusion. The enzyme chosen is AK3L1, an isoform of adenylate kinase. To investigate its dynamics, we have carried out three single point mutations (A74C, A132C, and A209C) with the cysteine residues being labeled with a coumarin-based solvatochromic probe [CPM: (7-diethylamino-3-(4-maleimido-phenyl)-4-methylcoumarin)]. Both enzyme activity and dynamics decreased in the binary mixtures as compared with the sum of the individual crowders, suggesting a reduction in excluded volume (in the mixture). To gain deeper insights into the binary mixtures, fluorescence correlation spectroscopy studies were carried out using fluorescein isothiocyanate-labeled Dextran 70 and tetramethylrhodaminelabeled AK3L1 as the diffusion probes. Diffusion in binary mixtures was observed to be much more constrained (relative to the sum of the individual crowders) for the labeled enzyme as compared to the labeled crowder showing different environments being faced by the two species. This was further confirmed during imaging of the phase-separated droplets formed in the binary mixtures having PEG as one of the crowding agents. The interior of these droplets was found to be rich in crowders and densely packed, as shown by confocal and digital holographic microscopy images, with the enzymes predominantly residing outside these droplets, that is, in the relatively less crowded regions. Taken together, our data provide important insights into various aspects of the simplest form of mixed crowding, that is, composed of just two components, and also hint at the enhanced complexity that the cellular interior presents toward having a detailed and comprehensive understanding of the same.

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“…Moreover, in vivo cells are not present as singular components rather they are present as a single layer of epithelial, endothelial or other stromal cells supported on a BM 26 . This is further augmented by a recent study from Ippolito et al which emphasizes that in vivo like mechanical and biological function of the tissue can only be attained by suitable morphological and cellular arrangement of cells in a monolayer 45 . This highlights the importance of scaffold design to achieve functional monolayers.…”

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confidence: 99%

Topographical influence of electrospun basement membrane mimics on formation of cellular monolayer

Jain

Rimal

Möller

et al. 2023

Sci Rep

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Functional unit of many organs like lung, kidney, intestine, and eye have their endothelial and epithelial monolayers physically separated by a specialized extracellular matrix called the basement membrane. The intricate and complex topography of this matrix influences cell function, behavior and overall homeostasis. In vitro barrier function replication of such organs requires mimicking of these native features on an artificial scaffold system. Apart from chemical and mechanical features, the choice of nano-scale topography of the artificial scaffold is integral, however its influence on monolayer barrier formation is unclear. Though studies have reported improved single cell adhesion and proliferation in presence of pores or pitted topology, corresponding influence on confluent monolayer formation is not well reported. In this work, basement membrane mimic with secondary topographical cues is developed and its influence on single cells and their monolayers is investigated. We show that single cells cultured on fibers with secondary cues form stronger focal adhesions and undergo increased proliferation. Counterintuitively, absence of secondary cues promoted stronger cell–cell interaction in endothelial monolayers and promoted formation of integral tight barriers in alveolar epithelial monolayers. Overall, this work highlights the importance of choice of scaffold topology to develop basement barrier function in in vitro models.

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The influence of entropic crowding in cell monolayers

Cited by 5 publications

References 42 publications

Tumor-Derived Membrane Vesicles Restrain Migration in Gliomas By Altering Collective Polarization

Tumor-Derived Membrane Vesicles Restrain Migration in Gliomas By Altering Collective Polarization

Deeper Insights into Mixed Crowding through Enzyme Activity, Dynamics, and Crowder Diffusion

Topographical influence of electrospun basement membrane mimics on formation of cellular monolayer

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