3D culture increases pluripotent gene expression in mesenchymal stem cells through relaxation of cytoskeleton tension

Zhou, Ying; Chen, Haiyan; Li, Hong; Wu, Yaojiong

doi:10.1111/jcmm.12946

Cited by 99 publications

(88 citation statements)

References 64 publications

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“…To investigate whether decreasing actin polymerization causes the reduction of MSC size, we examined the influence of 3D cell culture, CytoD treatment, and CD29 blockade on actin polymerization and cell size. Consistent with our previous findings , 3D culture dramatically reduced the amount of F‐actin filament bundles in the cytoplasm in immunofluorescence analysis (Fig. A) and the levels of total actin and F‐actin as determined by Western blot (Fig.…”

Section: Resultssupporting

confidence: 92%

“…Besides the reduction in cell size, we found that 3D spheroid culture dramatically decreased the expression of β1 integrin (CD29) along with several α intergrin binding chains in MSCs . In addition, the cells exhibited a marked reduction in the amount of actin filament bundles . Previous studies suggest that actin cytoskeleton play a role in regulating cell size; actin polymerization inhibits microparticle shedding and disruption of the cortical actin network increases the number of microparticles generated by leukocytes .…”

Section: Introductionmentioning

confidence: 73%

“…We and others have shown previously that three‐dimensional (3D) culture of MSCs in spheres for 36–60 hours dramatically reduces the cell volume by up to 70% . Associated with the reduction of cell size is substantially decreased entrapment of MSCs in the lungs after intravenous infusion and a markedly increased levels of circulating MSCs , suggesting an increased trafficking ability of the cells.…”

Section: Introductionmentioning

confidence: 89%

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Three-Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion

Zhou

et al. 2017

Stem Cells

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Our previous study has shown that three-dimensional (3D) culture decreases mesenchymal stem cell (MSC) size, leading to enhanced trafficking ability and reduced lung vascular obstructions. However, the underlying mechanisms are unclear. In this study, we proposed that 3D culture reduces MSC size by increasing vesicle excretion. Scanning electron microscope showed that 3D culture markedly increased the amount of membrane-bound vesicles on the cell surface. In consistence, tunable resistive pulse sensing quantifying analysis of vesicles in the culture medium indicated that there were higher levels of vesicles in the 3D culture MSC medium. 3D culture significantly lowered the level of actin polymerization (F-actin), suggestive of lowering actin skeleton tension may facilitate vesicle excretion. Indeed, treatment of MSCs with Cytochalasin D or functional blockade of integrin β1 caused increased vesicle secretion and decreased cell sizes. Thus, our results suggest that 3D culture reduces MSC size by increasing vesicle excretion which is likely mediated by lowering cytoskeleton tension. Stem Cells 2018;36:286-292.

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Section: Resultssupporting

confidence: 92%

Section: Introductionmentioning

confidence: 73%

Section: Introductionmentioning

confidence: 89%

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Three-Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion

Zhou

et al. 2017

Stem Cells

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“…The increased COX-2 expression is in turn due to the unbundled form of F-actin (i.e. a more relaxed form of actin, in comparison to the dense stress fibers observed in 2D) near the edge of the MBs, which was reported to sustain NF-κB activity 50 and to downregulate COX-2 transcriptional repressors 51,52 . Therefore NF-κB has a high activity in the outer layers of the MB, where it locally promotes COX-2 expression.…”

Section: Discussionmentioning

confidence: 98%

Mapping Structure and Biological Functions within Mesenchymal Bodies using Microfluidics

Sart

Tomasi

Barizien

et al. 2019

Preprint

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Organoids that recapitulate the functional hallmarks of anatomic structures comprise cell populations able to self-organize cohesively in 3D. However, the rules underlying organoid formation in vitro remain poorly understood because a correlative analysis of individual cell fate and spatial organization has been challenging. Here, we use a novel microfluidics platform to investigate the mechanisms determining the formation of organoids by human mesenchymal stromal cells that recapitulate the early steps of condensation initiating bone repair in vivo. We find that heterogeneous mesenchymal stromal cells selforganize in 3D in a developmentally hierarchical manner. We demonstrate a link between structural organization and local regulation of specific molecular signaling pathways such as NF-κB and actin polymerization, which modulate osteo-endocrine functions. This study emphasizes the importance of resolving spatial heterogeneities within cellular aggregates to link organization and functional properties, enabling a better understanding of the mechanisms controlling organoid formation, relevant to organogenesis and tissue repair.! 3!

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“…For example, human placenta MSC spheroids displayed lower expression of integrin β1 compared to normal 2D culture, leading to a decrease of cell-ECM interaction and enhancement of MSC stemness. By other approaches to release cytoskeleton tension, the stemness of MSCs was promoted as well, revealing the negative relationship between stemness and cytoskeleton tension in MSCs [71]. In addition to cell-matrix interaction, the cadherin-mediated cell-cell interaction cannot be ignored.…”

Section: Physical and Chemical Factors Affecting Cell Behaviors In Scmentioning

confidence: 99%

Cellular Spheroids of Mesenchymal Stem Cells and Their Perspectives in Future Healthcare

2019

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Intrinsic cellular properties of several types of cells are dramatically altered as the culture condition shifts from two-dimensional (2D) to three-dimensional (3D) environment. Currently, several lines of evidence have demonstrated the therapeutic potential of mesenchymal stem cells (MSCs) in regenerative medicine. MSCs not only replenish the lost cells, they also promote the regeneration of impaired tissues by modulating the immune responses. Following the development of 3D cell culture, the enhanced therapeutic efficacy of spheroid-forming MSCs have been identified in several animal disease models by promoting differentiation or trophic factor secretion, as compared to planar-cultured MSCs. Due to the complicated and multifunctional applications in the medical field, MSCs are recently named as medicinal signaling cells. In this review, we summarize the predominant differences of cell-environment interactions for the MSC spheroids formed by chitosan-based substrates and other scaffold-free approaches. Furthermore, several important physical and chemical factors affecting cell behaviors in the cell spheroids are discussed. Currently, the understanding of MSCs spheroid interactions is continuously expanding. Overall, this article aims to review the broad advantages and perspectives of MSC spheroids in regenerative medicine and in future healthcare. Three-Dimensional (3D) Cell Culture SystemsCell-cell and cell-environment interactions cooperatively determine the physiological phenomena in vivo. Many cell growth mechanisms and signal transduction pathways occurring in cells have been unveiled by the traditional two-dimensional (2D) culture system. Plastic-based culture materials remain the predominant cell culture platforms to date, yet some concerns have been raised with the accumulating experimental evidence.A major drawback of cell culture on 2D plastic plates is the planar cell-cell interaction that has the insufficient and inappropriate cell-environment crosstalk. In vivo, cells are grown and stacked within a three-dimensional (3D) space, and extracellular matrix (ECM) secreted by cells constitutes the supporting framework to further organize tissues and organs. Therefore, some important cell-cell and cell-environment interactions may be ignored in a planar culture platform. Primary cells, cancer cells, and stem cells have been cultured in a 3D environment, and many interesting findings have been uncovered. In general, the definition of 3D cell culture systems is that cells are cultured on/in the 3D scaffolds composed by natural or synthetic materials, or they are organized into cellular spheroids. With the continuing development of 3D cell culture, it is now generally agreed that the cell physiology and cell behavior are dramatically distinct for cells cultured within a 2D or 3D environment [1][2][3].Cancer cells are the most common cell types that have been investigated by 3D culture systems, and obvious change of gene expression as well as intracellular signaling in these 3D-cultured cancer cells have been ...

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3D culture increases pluripotent gene expression in mesenchymal stem cells through relaxation of cytoskeleton tension

Cited by 99 publications

References 64 publications

Three-Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion

Three-Dimensional Culture Reduces Cell Size By Increasing Vesicle Excretion

Mapping Structure and Biological Functions within Mesenchymal Bodies using Microfluidics

Cellular Spheroids of Mesenchymal Stem Cells and Their Perspectives in Future Healthcare

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