Growth factor and macromolecular crowding supplementation in human tenocyte culture

Tsiapalis, Dimitrios; Kearns, Stephen; Kelly, Jack L.; Zeugolis, Dimitrios I.

doi:10.1016/j.bbiosy.2021.100009

Cited by 15 publications

(11 citation statements)

References 133 publications

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“…Recent studies have argued that MMC may enable the accelerated development of functional tissue engineered medicines [ [34] , [35] , [36] ]. Indeed, in both hTC [ [37] , [38] , [39] , [40] ] and hDF [ [41] , [42] , [43] ] cultures, MMC has been shown to enhance (up to 120-fold) and accelerate (from 2 to 6 days) ECM deposition and to maintain hTC phenotype alone or in combination with other in vitro microenvironment modulators. It is also worth noting that the therapeutic potential of MMC has been demonstrated in preclinical setting [ 44 ], paving the way for its clinical translation.…”

Section: Introductionmentioning

confidence: 99%

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

Djalali-Cuevas,

Rettel,

Stein

et al. 2024

Materials Today Bio

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Section: Introductionmentioning

confidence: 99%

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

Djalali-Cuevas,

Rettel,

Stein

et al. 2024

Materials Today Bio

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“…We attribute this to the synergistic effect of the contained growth factors with MMC. Indeed, MMC combined with growth factor supplementation resulted in amplified (over cells with growth factors alone and cells with MMC alone) collagen deposition, as the growth factors enhanced collagen synthesis and MMC enhanced collagen deposition ( Tsiapalis et al, 2021 ). Following the same reasoning, one would have expected the ACF with MMC to induce higher collagen deposition than the ACF alone, which was not the case, as judged by SDS-PAGE.…”

Section: Discussionmentioning

confidence: 99%

“…Similarly, the presence of tissue-specific ECM has been shown to facilitate cell phenotype maintenance in vitro ( Pei et al, 2011 ; Cheng et al, 2014 ; Gattazzo et al, 2014 ; Yang et al, 2018a ). In eukaryotic cell culture systems, macromolecular crowding (MMC), following the principles of excluded volume effect, enhances and accelerates tissue-specific ECM deposition ( Raghunath and Zeugolis, 2021 ; Tsiapalis and Zeugolis, 2021 ; Zeugolis, 2021 ), a phenomenon that has been well documented in both differentiated ( Lareu et al, 2007 ; Satyam et al, 2014 ; Kumar et al, 2015a ; Kumar et al, 2015b ; Satyam et al, 2016 ; Kumar et al, 2018 ; Gaspar et al, 2019 ; Shendi et al, 2019 ; Tsiapalis et al, 2021 ) and progenitor cell cultures ( Zeiger et al, 2012 ; Prewitz et al, 2015 ; Cigognini et al, 2016 ; Lee et al, 2016 ; Patrikoski et al, 2017 ; Graceffa and Zeugolis, 2019 ; De Pieri et al, 2020 ). However, to-date, only one study has assessed the influence of MMC in xeno-free and/or serum-free media formulations using human adipose-derived mesenchymal stromal cells ( Patrikoski et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Macromolecular crowding in animal component-free, xeno-free and foetal bovine serum media for human bone marrow mesenchymal stromal cell expansion and differentiation

Korntner

Nubila²,

Gaspar³

et al. 2023

Front. Bioeng. Biotechnol.

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Background: Cell culture media containing undefined animal-derived components and prolonged in vitro culture periods in the absence of native extracellular matrix result in phenotypic drift of human bone marrow stromal cells (hBMSCs).Methods: Herein, we assessed whether animal component-free (ACF) or xeno-free (XF) media formulations maintain hBMSC phenotypic characteristics more effectively than foetal bovine serum (FBS)-based media. In addition, we assessed whether tissue-specific extracellular matrix, induced via macromolecular crowding (MMC) during expansion and/or differentiation, can more tightly control hBMSC fate.Results: Cells expanded in animal component-free media showed overall the highest phenotype maintenance, as judged by cluster of differentiation expression analysis. Contrary to FBS media, ACF and XF media increased cellularity over time in culture, as measured by total DNA concentration. While MMC with Ficoll™ increased collagen deposition of cells in FBS media, FBS media induced significantly lower collagen synthesis and/or deposition than the ACF and XF media. Cells expanded in FBS media showed higher adipogenic differentiation than ACF and XF media, which was augmented by MMC with Ficoll™ during expansion. Similarly, Ficoll™ crowding also increased chondrogenic differentiation. Of note, donor-to-donor variability was observed for collagen type I deposition and trilineage differentiation capacity of hBMSCs.Conclusion: Collectively, our data indicate that appropriate screening of donors, media and supplements, in this case MMC agent, should be conducted for the development of clinically relevant hBMSC medicines.

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“…This is much lower than that in vivo, and can cause cells to undergo epigenetic changes, leading to altered protein and gene pro les, phenotypic drift, loss of function and senescence. As a result, this can't provide an ideal cellular model for vascular diseases research such as hypertension (7,14). Adding inert polydisperse macromolecules to the culture medium can simulate the in vivo crowding environment (15).…”

Section: Introductionmentioning

confidence: 99%

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.

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Growth factor and macromolecular crowding supplementation in human tenocyte culture

Cited by 15 publications

References 133 publications

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

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

Macromolecular crowding in animal component-free, xeno-free and foetal bovine serum media for human bone marrow mesenchymal stromal cell expansion and differentiation

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

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