Study of cell behaviour on a cellulose anti-adhesive substratum

Velzenberger, Elodie; Vayssade, Muriel; Legeay, G.; Nagel, Marie‐Danielle

doi:10.1007/s10570-007-9182-4

Cited by 17 publications

(16 citation statements)

References 52 publications

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“…Cell proliferation assays showed that B16F10 cells grow more slowly in 3D culture on polyHEMA, whereas Swiss 3T3 cells are unable to grow in similar experimental conditions. We have previously shown that the proliferation index of B16F10 cells cultured for 48 h on polyHEMA is significantly lower than that of the same cells on tPS (Velzenberger et al, 2008). The present study indicates that melanoma cells can proliferate for 3 days in 3D architecture on polyHEMA, and only after that do cell numbers decline.…”

Section: B16f10 Cell Behaviour In 3d Culture On Polyhemasupporting

confidence: 49%

“…The adhesive proteins, like fibronectin and vitronectin, present in the serum‐enriched culture medium are immediately adsorbed onto the surface, so allowing cells to adhere to the ECM. Hence, cells initially respond to the adsorbed proteins, rather than to the surface itself (Shin et al, 2003; Wilson et al, 2005; Pakstis et al, 2009; Zhu et al, 2009). In cell–ECM adhesions, clustered integrin receptors bind to the ECM and connect to the actin cytoskeleton.…”

Section: Introductionmentioning

confidence: 99%

“…Our previous studies have focused on the highly metastatic B16F10 mouse melanoma cells, which spread and grow well on tPS, and adopt a 3D architecture in aggregates on anti-adhesive cellulose or polyHEMA [poly(2-hydroxyethylmethacrylate)]-coated polystyrene (Hindie et al, 2005(Hindie et al, , 2006Velzenberger et al, 2008). The aggregated B16F10 cells produce large amounts of a5 integrin, and the clusters of a5 integrin are co-localized with secreted fibronectin at the aggregate membranes.…”

Section: Introductionmentioning

confidence: 99%

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Growth and survival signalling in B16F10 melanoma cells in 3D culture

Goundiam

Nagel

Vayssade

2010

Cell Biology International

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The two-way communication between the ECM (extracellular matrix) and the cytoplasm via the integrins has many functions in cancer cells, including the suppression of apoptosis. As cells in a 3D (three-dimensional) architecture resemble the in vivo situation more closely than do cells in more conventional 2D cultures, we have employed a substratum that prevents cell adhesion and induces cell aggregation to determine why highly metastatic B16F10 melanoma cells resist anoikis. We compared the behaviour of B16F10 cells in 2D [on tPS (tissue culture polystyrene)] and 3D culture {on polyHEMA [poly(2-hydroxyethylmethacrylate)]} configurations. For this, we analysed cell morphology, proliferation, apoptosis and the activation status of several proteins involved in cell proliferation and survival [RhoA, FAK (focal adhesion kinase), Akt, ERK1/2 (extracellular-signal-regulated kinase 1/2)]. B16F10 cells in 3D architecture were able to proliferate as cell aggregates for 3 days, after which the number of cells decreased. The normal Swiss 3T3 cells used as an anoikis-sensitive control did not proliferate on the anti-adhesive substratum. Rho A was activated in B16F10 aggregates throughout their time in culture, whereas it was not in Swiss 3T3 aggregates. An absence of apoptotic activity was correlated with the proliferation of B16F10 cells in aggregates: caspase 3 was significantly activated only after 3 days in culture on polyHEMA. FAK and Akt were transiently activated, and their inactivation was correlated with the induction of apoptosis. ERK1/2 were activated throughout the 3D culture. No survival protein was activated in Swiss 3T3 aggregates. Data obtained from cells in 3D culture suggest that B16F10 cells are resistant to anoikis through the activation of the FAK and Akt signalling pathways.

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Section: B16f10 Cell Behaviour In 3d Culture On Polyhemasupporting

confidence: 49%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Growth and survival signalling in B16F10 melanoma cells in 3D culture

Goundiam

Nagel

Vayssade

2010

Cell Biology International

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“…B16F10 melanoma cells seeded on tPS were well spread, with a well‐organized actin cytoskeleton and stress fibre formation, whereas cells cultured on a polyHEMA substratum formed large aggregates, with no well‐organized actin cytoskeleton (Goundiam et al, 2010). Most B16F10 cells were in G 1 phase, but they were still capable of growing for 3 days, whereas ‘normal’ Swiss 3T3 cells used as anoikis‐sensitive control did not proliferate (Velzenberger et al, 2008; Goundiam et al, 2010). After 3 days, B16F10 cells underwent caspase‐3‐dependent apoptosis.…”

Section: Discussionmentioning

confidence: 99%

Akt and RhoA inhibition promotes anoikis of aggregated B16F10 melanoma cells

Goundiam

Nagel

Vayssade

2012

Cell Biology International

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In the highly metastatic B16F10 melanoma cell line, activation of the signalling molecules that promote cell proliferation and survival on conventional adhesive culture dishes may also be responsible for the growth and resistance to anoikis of aggregates on a non-adhesive substratum. We have examined the influence of bacterial ADP-ribosyltransferases C3-like exoenzymes, which selectively modify RhoA, B and C proteins and inhibit signal pathways controlled by them. RNA interference [siRNA (small interfering RNA) Akt (also known as protein kinase B)] and a PI3K (phosphoinositide 3-kinase) inhibitor were used to analyse the changes caused by inhibiting the PI3K/Akt pathway. Inhibiting the activation of RhoA, B, C and Akt expression resulted in a decrease of the number of cells cultured in aggregates, and caspase 3 activation. RhoA activation and RhoB and RhoC expression were controlled by Akt, but not RhoA expression. Inhibiting Akt and RhoA reduced the expression of α5 integrin, and inactivated FAK (focal adhesion kinase) in B16F10 cells cultured as aggregates. Thus, inhibiting Rho subfamily proteins and Akt expression inactivates the FAK pathway and induces anoikis in anoikis-resistant cells. The activation of RhoA in melanoma cells can depend on PI3K/Akt activation, suggesting that PI3K/Akt is a suitable target for new therapeutic approaches.

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“…Additionally, cellulose originating from plants is the most abundant natural polymer on earth. Therefore, cellulose has been used as a matrix material for cell culture in several studies, in particular cellulose derivatives such as hydroxypropylmethylcellulose and carboxymethylcellulose [15][16][17][18], but also bacterial cellulose which had been made cell-adhesive by chemical treatment [19,20]. Currently, there are few studies reporting on rat hepatocytes.…”

Section: Introductionmentioning

confidence: 99%

Microfibrillated Cellulose Sheets Coating Oxygen-Permeable PDMS Membranes Induce Rat Hepatocytes 3D Aggregation into Stably-Attached 3D Hemispheroids

Evenou

Couderc

Kim

et al. 2011

Journal of Biomaterials Science, Polymer Edition

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Here we report the use of natural, chemically-unmodified, microfibrillated cellulose (MFC) as a matrix for hepatocyte culture. We developed an original cell-culture design composed of a thin 3D-microstructured fibrous substrate consisting of a MFC sheet coating a highly O(2)-permeable polydimethylsiloxane (PDMS) membrane. The MFC-coated PDMS membranes were obtained according to a simple process where cellulose fibres were deposited from an aqueous suspension on the PDMS surfaces and the films were dried under mild conditions. To enable oxygen diffusion through the membranes, they were assembled on bottomless frames ('O(2)+' condition). Rat hepatocytes primary-cultured on such MFC-PDMS membranes quickly organized themselves into large hemispherical 3D aggregates which were tightly anchored to the MFC sheets. In contrast, hepatocytes cultured on smooth PDMS membranes in the O(2)+ system (O(2)+, PDMS) organized into unstable 2D monolayers which easily detached from the surfaces. Hepatocyte 3D cultures obtained on MFC-PDMS membranes exhibited higher liver-specific functions over a 2-week culture period, as assessed by both the higher albumin secretion and urea synthesis rate. The MFC-PDMS membranes appear suitable for obtaining stably-attached and functional hepatocyte 3D cultures and appear interesting for drug/chemical screenings in a microplate format, but also for microfluidic applications.

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Study of cell behaviour on a cellulose anti-adhesive substratum

Cited by 17 publications

References 52 publications

Growth and survival signalling in B16F10 melanoma cells in 3D culture

Growth and survival signalling in B16F10 melanoma cells in 3D culture

Akt and RhoA inhibition promotes anoikis of aggregated B16F10 melanoma cells

Microfibrillated Cellulose Sheets Coating Oxygen-Permeable PDMS Membranes Induce Rat Hepatocytes 3D Aggregation into Stably-Attached 3D Hemispheroids

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