Direct Observation of Tunneling Nanotubes within Human Mesenchymal Stem Cell Spheroids

Zhang, Jiali; Whitehead, Jacklyn; Liu, Yang; Yang, Qingbo; Leach, J. Kent; Liu, Gang-yu

doi:10.1021/acs.jpcb.8b07305

Cited by 20 publications

(24 citation statements)

References 34 publications

(75 reference statements)

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“…We observed decreases in VEGF ( Figure 2A other reports identifying TNTs. 18,28 TNTs are actin-rich structures that can be inhibited in the presence of CytoD. 18 To ensure that CytoD treatment did not impair MSC spheroid health, we evaluated the viability, total DNA content, and metabolic activity of spheroids within each group.…”

Section: Resultsmentioning

confidence: 99%

“…To block TNT formation, cytochalasin D (CytoD, 500 nM, Sigma-Aldrich, St. Louis, Missouri) was added to each well immediately after centrifugation, and cells were exposed for the full 48 hour incubation. 18 The diameter of live spheroids was measured using brightfield microscopy (Nikon Eclipse TE2000U microscope, Melville, New York).…”

Section: Spheroid Formation and Characterizationmentioning

confidence: 99%

“…This suspension was pipetted onto poly-L-lysine-coated silicon wafers, followed by a multi-step dehydration process before being placed inside the SEM chamber. 18 Spheroids were imaged using a field emission SEM (S-4100T FE-SEM, Hitachi High Technologies America, Pleasanton, California) under an accelerating voltage of 2 kV at 10 μA.…”

Section: Scanning Electron Microscopy Imagingmentioning

confidence: 99%

“…Prior to spheroid formation, MSCs were stained with 5 μM of CellTrace™ Far Red and/or Violet (Thermo Fisher) following the manufacturer's recommendations. 18…”

Section: Quantification Of Cytosolic Colocalization Using Laser Scamentioning

confidence: 99%

“…16 TNTs have been reported in 3D aggregates of cancerous cells 17 and more recently in MSC spheroids by our group. 18 These intercellular bridges shuttle cytoplasmic content and support distressed cells. 19,20 MSCs possess protective and restorative potential in co-culture systems that may be partially mediated by TNT formation between cells, 16 providing an opportunity to overcome the limitations of acquired senescence.…”

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confidence: 99%

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Tunneling nanotubes mediate the expression of senescence markers in mesenchymal stem/stromal cell spheroids

et al. 2019

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The therapeutic potential of mesenchymal stem/stromal cells (MSCs) is limited by acquired senescence following prolonged culture expansion and high-passage numbers.However, the degree of cell senescence is dynamic, and cell-cell communication is critical to promote cell survival. MSC spheroids exhibit improved viability compared with monodispersed cells, and actin-rich tunneling nanotubes (TNTs) may mediate cell survival and other functions through the exchange of cytoplasmic components. Building upon our previous demonstration of TNTs bridging MSCs within these cell aggregates, we hypothesized that TNTs would influence the expression of senescence markers in MSC spheroids. We confirmed the existence of functional TNTs in MSC spheroids formed from low-passage, high-passage, and mixtures of low-and high-passage cells using scanning electron microscopy, confocal microscopy, and flow cytometry. The contribution of TNTs toward the expression of senescence markers was investigated by blocking TNT formation with cytochalasin D (CytoD), an inhibitor of actin polymerization. CytoD-treated spheroids exhibited decreases in cytosol transfer. Compared with spheroids formed solely of high-passage MSCs, the addition of low-passage MSCs reduced p16 expression, a known genetic marker of senescence. We observed a significant increase in p16 expression in high-passage cells when TNT formation was inhibited, establishing the importance of TNTs in MSC spheroids. These data confirm the restorative role of TNTs within MSC spheroids formed with low-and high-passage cells and represent an exciting approach to use higher-passage cells in cell-based therapies. K E Y W O R D Smesenchymal stem/stromal cells, p16, senescence, spheroids, tunneling nanotubes

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Section: Resultsmentioning

confidence: 99%

Section: Spheroid Formation and Characterizationmentioning

confidence: 99%

Section: Scanning Electron Microscopy Imagingmentioning

confidence: 99%

“…Prior to spheroid formation, MSCs were stained with 5 μM of CellTrace™ Far Red and/or Violet (Thermo Fisher) following the manufacturer's recommendations. 18…”

Section: Quantification Of Cytosolic Colocalization Using Laser Scamentioning

confidence: 99%

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confidence: 99%

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Tunneling nanotubes mediate the expression of senescence markers in mesenchymal stem/stromal cell spheroids

et al. 2019

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Harnessing mesenchymal aggregation for engineered organ‐level regeneration: Recent progress and perspective

Zheng,

Sui,

Jin

2023

Aggregate

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Stem cells, especially mesenchymal progenitors or mesenchymal stem cells (MSCs), possess an intrinsic property to form compact spheroid‐like assemblies, a phenomenon known as cell aggregation. In recent years, a growing body of researches have uncovered that this is a cross‐species conserved developmental event essential for initiating organogenesis in a variety of organs. Moreover, the self‐assembly property also contributes to the regenerative capacities of MSC aggregates in vivo with broad range of applications in tissue engineering. In this review, the principles of self‐assembled mesenchymal aggregation and its involvement in physiological organogenesis, as well as the construction approaches of engineering MSC aggregates and its application for organ regeneration are discussed. The authors aim to provide a speculative overview of the current understanding and the recent findings of cell aggregation, from both the developmental and the engineering perspectives, and thus offer insights into the understanding of stem cell biology and the establishment of novel organ regeneration strategies.

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Tunneling nanotubes: A bridge for heterogeneity in glioblastoma and a new therapeutic target?

Venkatesh

Lou

2019

Cancer Reports

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BackgroundThe concept of tumour heterogeneity is not novel but is fast becoming a paradigm by which to explain part of the highly recalcitrant nature of aggressive malignant tumours. Glioblastoma is a prime example of such difficult‐to‐treat, invasive, and incurable malignancies. With the advent of the post‐genomic age and increased access to next‐generation sequencing technologies, numerous publications have described the presence and extent of intratumoural and intertumoural heterogeneity present in glioblastoma. Moreover, there have been numerous reports more directly correlating the heterogeneity of glioblastoma to its refractory, reoccurring, and inevitably terminal nature. It is therefore prudent to consider the different forms of heterogeneity seen in glioblastoma and how to harness this understanding to better strategize novel therapeutic approaches. One of the most central questions of tumour heterogeneity is how these numerous different cell types (both tumour and non‐tumour) in the tumour mass communicate.Recent findingsThis chapter provides a brief review on the variable heterogeneity of glioblastoma, with a focus on cellular heterogeneity and on modalities of communication that can induce further molecular diversity within the complex and ever‐evolving tumour microenvironment. We provide particular emphasis on the emerging role of actin‐based cellular conduits called tunnelling nanotubes (TNTs) and tumour microtubes (TMs) and outline the perceived current problems in the field that need to be resolved before pharmacological targeting of TNTs can become a reality.ConclusionsWe conclude that TNTs and TMs provide a new and exciting avenue for the therapeutic targeting of glioblastoma and that numerous inroads have already made into TNT and TM biology. However, to target TMs and TNTs, several advances must be made before this aim can become a reality.

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Direct Observation of Tunneling Nanotubes within Human Mesenchymal Stem Cell Spheroids

Cited by 20 publications

References 34 publications

Tunneling nanotubes mediate the expression of senescence markers in mesenchymal stem/stromal cell spheroids

Tunneling nanotubes mediate the expression of senescence markers in mesenchymal stem/stromal cell spheroids

Harnessing mesenchymal aggregation for engineered organ‐level regeneration: Recent progress and perspective

Tunneling nanotubes: A bridge for heterogeneity in glioblastoma and a new therapeutic target?

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