Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures

David, M; Kelly, Elizabeth; Zoellner, Hans

doi:10.1016/j.cyto.2013.02.028

Cited by 11 publications

(32 citation statements)

References 13 publications

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“…We earlier described the exchange of membrane and cytoplasmic protein between cultured human fibroblasts (Fib) and malignant cells (MC) (1). Others have made similar observations, and describe this as via either tunneling nanotubes (TNT) or exosomes and other shed membrane vesicles, and this is often associated with changes in cell phenotype (2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17), while we have also seen phenotypic change (1,18). Mitochondrial exchange has been considered particularly interesting (2,4,5,19).…”

Section: Introductionsupporting

confidence: 62%

Potential hydrodynamic cytoplasmic transfer between mammalian cells: Cell-projection pumping

Zoellner

Paknejad

Cornwell

et al. 2019

Preprint

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We earlier reported cytoplasmic fluorescence exchange between cultured human fibroblasts (Fib) and malignant cells (MC). Others report similar transfer via either tunneling nanotubes (TNT) or shed membrane vesicles and this changes the phenotype of recipient cells. Our current time-lapse microscopy showed most exchange was from Fib into MC, with less in the reverse direction. Although TNT were seen, we were surprised transfer was not via TNT, but was instead via fine and often branching cell projections that defied direct visual resolution because of their size and rapid movement. Their structure was revealed nonetheless, by their organellar cargo and the grooves they formed indenting MC. Discrete, rapid and highly localized transfer events, evidenced against a role for shed vesicles. Transfer coincided with rapid retraction of the cell-projections, suggesting a hydrodynamic mechanism. Increased hydrodynamic pressure in retracting cell-projections normally returns cytoplasm to the cell body. We hypothesize 'cellprojection pumping' (CPP), where cytoplasm in retracting cell-projections partially equilibrates into adjacent recipient cells via micro-fusions that form temporary inter-cellular cytoplasmic continuities. We tested plausibility for CPP by combined mathematical modelling, comparison of predictions from the model with experimental results, and then computer simulations based on experimental data. The mathematical model predicted preferential CPP into cells with lower cell stiffness, expected from equilibration of pressure towards least resistance. Predictions from the model were satisfied when Fib were cocultured with MC, and fluorescence exchange related with cell stiffness by atomic force microscopy. When transfer into 5000 simulated recipient MC or Fib was studied in computer simulations, inputting experimental cell stiffness and donor cell fluorescence values generated transfers to simulated recipient cells similar to those seen by experiment. We believe CPP is a novel mechanism in mammalian inter-cellular cytoplasmic transfer and communication.

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Section: Introductionsupporting

confidence: 62%

Potential hydrodynamic cytoplasmic transfer between mammalian cells: Cell-projection pumping

Zoellner

Paknejad

Cornwell

et al. 2019

Preprint

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“…Abrogation of cytokine activity by boiling supports the activity as dependent on native TNF-α conformation, and independence from Polymyxin-B is consistent with the absence of any effect by potentially contaminant lipopolysaccharide. While TNF-α and other ligands of the TNF family receptors can induce apoptosis in fibroblasts and other cells, this is typically only when protein and or mRNA synthesis are inhibited [27]–[31], while there was no sign of TNF-α induced apoptosis in either the current study or our earlier published work [9], [14].…”

Section: Discussionmentioning

confidence: 45%

“…However, where neoplastic cells invading from uninflamed tumour areas encounter local inflammation, cytokine stimulated fibroblasts may have more intense interaction with the malignant cells due to increased adhesion. While it is acknowledged that this scenario is highly speculative and would require in-vivo confirmation as well as further characterization of the SAOS-2 response to TNF-α, current data together with our separate report of altered cytokine synthesis by h-GF permitted cellular sipping [14], do highlight the extreme complexity of interactions between neoplastic and stromal cells and suggest at least partial explanation for the great morphological variability seen within different domains of single neoplasms in-vivo [38]–[40].…”

Section: Discussionmentioning

confidence: 73%

“…Expression of mRNA for the inflammatory cytokine Tumour Necrosis Factor-α (TNF-αin malignant and stromal cells is associated with poor prognosis [10], [11], and fibroblasts respond to this cytokine with increased adhesion molecule expression and malignant cell binding [12], [13], hence we also investigated the effect of TNF-α and found that this cytokine significantly increased cellular sipping between h-GF and SAOS-2 [9]. In separate work, we demonstrated altered cytokine synthesis in response to TNF-α by h-GF permitted cellular sipping, compared with h-GF denied this contact dependent interaction [14].…”

Section: Introductionmentioning

confidence: 88%

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SAOS-2 Osteosarcoma Cells Bind Fibroblasts via ICAM-1 and This Is Increased by Tumour Necrosis Factor-α

et al. 2014

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We recently reported exchange of membrane and cytoplasmic markers between SAOS-2 osteosarcoma cells and human gingival fibroblasts (h-GF) without comparable exchange of nuclear markers, while similar h-GF exchange was seen for melanoma and ovarian carcinoma cells. This process of “cellular sipping” changes phenotype such that cells sharing markers of both SAOS-2 and h-GF have morphology intermediate to that of either cell population cultured alone, evidencing increased tumour cell diversity without genetic change. TNF-α increases cellular sipping between h-GF and SAOS-2, and we here study binding of SAOS-2 to TNF-α treated h-GF to determine if increased cellular sipping can be accounted for by cytokine stimulated SAOS-2 binding. More SAOS-2 bound h-GF pe-seeded wells than culture plastic alone (p<0.001), and this was increased by h-GF pre-treatment with TNF-α (p<0.001). TNF-α stimulated binding was dose dependent and maximal at 1.16nM (p<0.05) with no activity below 0.006 nM. SAOS-2 binding to h-GF was independent of serum, while the lipopolysaccharide antagonist Polymyxin B did not affect results, and TNF-α activity was lost on boiling. h-GF binding of SAOS-2 started to increase after 30min TNF-α stimulation and was maximal by 1.5hr pre-treatment (p<0.001). h-GF retained maximal binding up to 6hrs after TNF-α stimulation, but this was lost by 18hrs (p<0.001). FACS analysis demonstrated increased ICAM-1 consistent with the time course of SAOS-2 binding, while antibody against ICAM-1 inhibited SAOS-2 adhesion (p<0.04). Pre-treating SAOS-2 with TNF-α reduced h-GF binding to background levels (p<0.003), and this opposite effect to h-GF cytokine stimulation suggests that the history of cytokine exposure of malignant cells migrating across different microenvironments can influence subsequent interactions with fibroblasts. Since cytokine stimulated binding was comparable in magnitude to earlier reported TNF-α stimulated cellular sipping, we conclude that TNF-α stimulated cellular sipping likely reflects increased SAOS-2 binding as opposed to enhanced exchange mechanisms.

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“…In the case of PPGP, however, the osteogenic differentiation was significantly enhanced when BMSCs were cultured directly on PPGP films in comparison with those cells being cultured in the non-contact manner. In our previous study and published data, [43][44][45] contact effect from the substrate on cell biological behaviors had been identified to be different from the effect of those soluble ingredients from the substrate, especially, when the substrate was bioactive. [44] In the present study, on one hand, the phosphorus-containing surface of polyphosphazene film was proven bioactive and osteocompatible.…”

Section: Comparative Analysis On Contact and Non-contact Culture Resultsmentioning

confidence: 99%

Molecular Mechanism Study on Effect of Biodegradable Amino Acid Ester–Substituted Polyphosphazenes in Stimulating Osteogenic Differentiation

Huang

Yang

et al. 2019

Macromolecular Bioscience

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Amino acid ester substituted polyphosphazenes are osteoactive benefiting from their phosphorus‐containing chemical structure, which highlights interests in bone tissue engineering. To correlate their chemical structures with cell activities, in this study, poly[(ethyl alanato)0.3(ethyl glycinato)0.7phosphazene] (PAGP) and poly[(ethyl phenylalanato)0.3(ethyl glycinato)0.7phosphazene] (PPGP) are synthesized to carry out studies on cell osteogenic differentiation. In the non‐contact culture manner, bone mesenchymal stromal cells (BMSCs) are cultured in transwell chambers containing PAGP or PPGP films, while the cells and the materials do not contact. In the contact culture manner, BMSCs are cultured on the PAGP or PPGP films. In the meantime, solutions containing PAGP or PPGP degradation products (i.e., phosphate, ammonium, and corresponding amino acids) are applied for cell culture using inorganic phosphate (Pi) ion as control. Thus, the influences from substrate surface and degradation products can be identified separately. The results reveal that both the phosphorus‐containing surface of PAGP and PPGP films and their degradation products play significant roles in regulating cell behaviors. In comparison with PAGP, PPGP seems able to provide relatively stable phosphorus‐containing surface to strengthen the cell‐scaffold interaction because of its slower degradation rate and higher Young's modulus, leading to greater promotion in osteogenic differentiation via contact effect.

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Opposite cytokine synthesis by fibroblasts in contact co-culture with osteosarcoma cells compared with transwell co-cultures

Cited by 11 publications

References 13 publications

Potential hydrodynamic cytoplasmic transfer between mammalian cells: Cell-projection pumping

Potential hydrodynamic cytoplasmic transfer between mammalian cells: Cell-projection pumping

SAOS-2 Osteosarcoma Cells Bind Fibroblasts via ICAM-1 and This Is Increased by Tumour Necrosis Factor-α

Molecular Mechanism Study on Effect of Biodegradable Amino Acid Ester–Substituted Polyphosphazenes in Stimulating Osteogenic Differentiation

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