Macromolecular crowding in human tenocyte and skin fibroblast cultures: A comparative analysis

Djalali-Cuevas, Adrian; Rettel, Mandy; Stein, Frank; Savitski, Mikhail; Kearns, Stephen; Kelly, Jack; Biggs, Manus; Skoufos, Ioannis; Tzora, Athina; Prassinos, Nikitas; Diakakis, Nikolaos; Zeugolis, Dimitrios I.

doi:10.1016/j.mtbio.2024.100977

Cited by 3 publications

(1 citation statement)

References 209 publications

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“…Thus, reducing the loss of ECM in the medium also needs further exploration [ 18 , 37 , 43 ]. Clusters 3 and 4 are intracellular organelles that regulate mitochondrial and cellular metabolism, RNA and amino acid biosynthesis, and metabolic pathways, consistent with the protein levels that had been reported in the literature [ 44 ]. This affects the overall rate of protein synthesis, which is a hallmark of cell and tissue growth.…”

Section: Discussionsupporting

confidence: 82%

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

Liu,

Jiang,

et al. 2024

Eur J Med Res

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Background The contractile phenotype of vascular smooth muscle cells (VSMCs) results in good diastolic and contractile capacities, and its altered function is the main pathophysiological basis for diseases such as hypertension. VSMCs exist as a synthetic phenotype in vitro, making it challenging to maintain a contractile phenotype for research. It is widely recognized that the common medium in vitro is significantly less crowded than in the in vivo environment. Additionally, VSMCs have a heightened sense for detecting changes in medium crowding. However, it is unclear whether macromolecular crowding (MMC) helps maintain the VSMCs contractile phenotype. Purpose This study aimed to explore the phenotypic, behavioral and gene expression changes of VSMCs after increasing the crowding degree by adding carrageenan (CR). Methods The degree of medium crowding was examined by a dynamic light scattering assay; VSMCs survival and activity were examined by calcein/PI cell activity and toxicity and CCK-8 assays; VSMCs phenotypes and migration were examined by WB and wound healing assays; and gene expression was examined by transcriptomic analysis and RT-qPCR. Results Notably, 225 μg/mL CR significantly increased the crowding degree of the medium and did not affect cell survival. Simultaneously, CR significantly promoted the contraction phenotypic marker expression in VSMCs, shortened cell length, decreased cell proliferation, and inhibited cell migration. CR significantly altered gene expression in VSMCs. Specifically, 856 genes were upregulated and 1207 genes were downregulated. These alterations primarily affect the cellular ion channel transport, microtubule movement, respiratory metabolism, amino acid transport, and extracellular matrix synthesis. The upregulated genes were primarily involved in the cytoskeleton and contraction processes of VSMCs, whereas the downregulated genes were mainly involved in extracellular matrix synthesis. Conclusions The in vitro study showed that VSMCs can maintain the contractile phenotype by sensing changes in the crowding of the culture environment, which can be maintained by adding CR. Graphical Abstract

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Section: Discussionsupporting

confidence: 82%

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

Liu,

Jiang,

et al. 2024

Eur J Med Res

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Crowd movement monitoring in academic buildings: a reinforcement learning approach

Geetha,

Rao,

Chellaswamy

2024

Multimed Tools Appl

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Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing Macromolecular crowding in vitro

Liu,

Jiang,

et al. 2024

Preprint

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Background The contractile phenotype of vascular smooth muscle cells (VSMCs) has a good diastolic and contractile capacity, and their altered function is the main pathophysiological basis for diseases such as hypertension. VSMCs always exist in a synthetic phenotype in vitro, making it challenging to maintain the contractile phenotype for research. It is widely recognized that common medium in vitro is significantly less crowded than the in vivo environment. Additionally, VSMCs have a heightened sense of touch to detect medium crowding changes. However, it is unclear whether macromolecular crowding (MMC) can help maintain the contractile phenotype of VSMCs. Purpose To study and obtain the phenotypic, behavioral and gene expression changes of VSMCs after increasing the crowding degree by adding carrageenan (CR). Methods The medium crowding degree was examined by the dynamic light scattering assay;VSMCs survival and activity were examined by the Calcein/PI cell activity and toxicity assay and CCK-8 assays༛VSMCs phenotype and migration were examined by WB and wound healing assay༛Gene expression was examined by transcriptomic analysis and RT-qPCR. Results 225 µg/mL CR significantly increased the crowding degree of the medium and did not affect cell survival. Simultaneously, CR significantly promoted contraction phenotypic markers expression in VSMCs, cell length was shortened, cell proliferation was decreased, and cell migration was inhibited. CR significantly altered gene expression in VSMCs. Specifically, 856 genes were up-regulated and 1207 genes were down-regulated. These alterations primarily affected cellular ion channel transport, microtubule movement, cellular respiratory metabolism, amino acid transport, and extracellular matrix synthesis. The up-regulated genes were primarily involved in the cytoskeleton and the contraction process of VSMCs, while the down-regulated genes were mainly involved in extracellular matrix synthesis. Conclusion The study showed that VSMCs in vitro can maintain the contractile phenotype by sensing changes in the culture environment's crowding, which can be maintained by adding a concentration of CR.

show abstract

Macromolecular crowding in human tenocyte and skin fibroblast cultures: A comparative analysis

Cited by 3 publications

References 209 publications

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing macromolecular crowding in vitro

Crowd movement monitoring in academic buildings: a reinforcement learning approach

Carrageenan maintains the contractile phenotype of vascular smooth muscle cells by increasing Macromolecular crowding in vitro

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