Cell proliferation in <i>in vivo</i>‐like three‐dimensional cell culture is regulated by sequestration of <scp>ERK</scp>1/2 to lipid rafts

Skrobanska, Ralica; Evangelatov, Aleksandar; Stefanova, Nadezhda; Topouzova-Hristova, Tanya; Momchilova, Albena; Pankov, Roumen

doi:10.1111/cpr.12112

Cited by 4 publications

(7 citation statements)

References 49 publications

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“…Cells on both (-) and (+)pST indeed showed lower growth rate than on flat pST, in parallel with occurrences in cells cultured in 3D models. 50,51 However, biophysical properties favored preferential adherence to (-)pST but not to (+)pST substrate, revealing that the details of topography were also important. This study is the first to develop a method in which the imprinted substrate has a cell's physical shapes but without differential cell secretion and surface chemistry complexities that are present in cell cluster models.…”

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confidence: 99%

The characteristics of Ishikawa endometrial cancer cells are modified by substrate topography with cell-like features and the polymer surface

Tan

Sykes

Alkaisi

et al. 2015

IJN

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Conventional in vitro culture studies on flat surfaces do not reproduce tissue environments, which have inherent topographical mechanical signals. To understand the impact of these mechanical signals better, we use a cell imprinting technique to replicate cell features onto hard polymer culture surfaces as an alternative platform for investigating biomechanical effects on cells; the high-resolution replication of cells offers the micro- and nanotopography experienced in typical cell–cell interactions. We call this platform a Bioimprint. Cells of an endometrial adenocarcinoma cell line, Ishikawa, were cultured on a bioimprinted substrate, in which Ishikawa cells were replicated on polymethacrylate (pMA) and polystyrene (pST), and compared to cells cultured on flat surfaces. Characteristics of cells, incorporating morphology and cell responses, including expression of adhesion-associated molecules and cell proliferation, were studied. In this project, we fabricated two different topographies for the cells to grow on: a negative imprint that creates cell-shaped hollows and a positive imprint that recreates the raised surface topography of a cell layer. We used two different substrate materials, pMA and pST. We observed that cells on imprinted substrates of both polymers, compared to cells on flat surfaces, exhibited higher expression of β1-integrin, focal adhesion kinase, and cytokeratin-18. Compared to cells on flat surfaces, cells were larger on imprinted pMA and more in number, whereas on pST-imprinted surfaces, cells were smaller and fewer than those on a flat pST surface. This method, which provided substrates in vitro with cell-like features, enabled the study of effects of topographies that are similar to those experienced by cells in vivo. The observations establish that such a physical environment has an effect on cancer cell behavior independent of the characteristics of the substrate. The results support the concept that the physical topography of a cell’s environment may modulate crucial oncological signaling pathways; this suggests the possibility of cancer therapies that target pathways associated with the response to mechanical stimuli.

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mentioning

confidence: 99%

The characteristics of Ishikawa endometrial cancer cells are modified by substrate topography with cell-like features and the polymer surface

Tan

Sykes

Alkaisi

et al. 2015

IJN

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“…To obtain naturally organized three-dimensional cell cultures we used GD25β1 cells that produce dense extracellular matrix and grow above confluency as multilayered tissue-like structures, where cells demonstrate reduced ageing and proliferation [12]. The general morphology of these 3D fibroblast cultures is illustrated after staining with hematoxylin/eosin in Figure 1.…”

Section: Resultsmentioning

confidence: 99%

“…Despite the numerous investigations devoted to the various effects of quercetin, the fine molecular mechanisms underlying its action on cells and membranes still need to be clarified. In this work we used as experimental model quercetin-treated three-dimensional tissue-like fibroblast cultures, which mimic closely the structure and functions of native tissues [12]. On this model we tried to reveal the biochemical basis of quercetin impact on cellular membranes, presuming that the obtained results would be more relevant to in vivo-like conditions than ones obtained on monolayer cells.…”

Section: Discussionmentioning

confidence: 99%

“…Cells were cultured at 37 °C, in a humidified air atmosphere supplemented with 5% CO 2 . Three-dimensional (3D) cell cultures were prepared as described by Skrobanska et al [12]. Briefly, the cells were seeded at a concentration of 0.67 × 10 5 cells per cm 2 and allowed to produce their own matrix by culturing them for five consecutive days under the conditions described above.…”

Section: Cell Culture and Histochemical Stainingmentioning

confidence: 99%

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Quercetin affects membrane lipids and apoptosis in three-dimensional fibroblast cultures

Momchilova

Markovska

Georgiev

et al. 2021

Biotechnology & Biotechnological Equipment

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The polyphenol quercetin is associated with numerous beneficial health effects in various pathologies such as oxidative stress, neurodegenerative disorders, inflammation, neoplastic processes, age-related diseases, and so on. However, the molecular mechanisms underlying the effects of quercetin and its implication in the pharmaceutical practice require additional clarification. In this study we analyzed the biochemical mechanisms of quercetin effect on membrane lipids of fibroblasts cultured as three-dimensional (3D) cultures, which resemble living tissues more adequately compared to cellular monolayers. Quercetin treatment of 3D fibroblasts induced alterations of the plasma membrane phospholipid and fatty acid composition as well as in the phospholipid metabolizing enzymes sphingomyelinase and phospholipase A 2 . The level of sphingomyelin (SM), which acts as endogenous membrane antioxidant, was elevated due to quercetin action. Incubation of 3D fibroblasts with quercetin induced augmentation in the saturated fatty acids and reduction of all polyunsaturated fatty acids in the membrane phospholipid molecules of the plasma membranes, indicating an additional increase of the resistance to oxidative damage. Quercetin treatment reduced cholesterol susceptibility to oxidation, a phenomenon which is probably related to the observed increase in the level of SM. In addition, Western blot analysis showed augmented phosphorylation of Akt and expression of Bcl-2 in quercetin-treated cell, indicating that apoptosis was suppressed by quercetin. In conclusion, the reported results contribute to a better understanding of the beneficial effects of quercetin on the structure and functions of plasma membranes of 3D cell cultures, which are a more adequate model of living tissues.

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“…Performing a 2D co-culture, using cells with different phenotypes, increases the similarity regarding the cellular heterogeneity observed in vivo models; however, this is still not enough when compared to living tissues [ 11 , 12 , 13 , 14 , 15 , 16 ]. In this sense, a 3D co-culture is a more promising strategy for creating integrated cross-communication networks in vitro [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Alginate-Based 3D A549 Cell Culture Model to Study Paracoccidioides Infection

Santos

Oliveira

Fontes

et al. 2023

JoF

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A three-dimensional (3D) lung aggregate model based on sodium alginate scaffolds was developed to study the interactions between Paracoccidioides brasiliensis (Pb) and lung epithelial cells. The suitability of the 3D aggregate as an infection model was examined using cell viability (cytotoxicity), metabolic activity, and proliferation assays. Several studies exemplify the similarity between 3D cell cultures and living organisms, which can generate complementary data due to the greater complexity observed in these designed models, compared to 2D cell cultures. A 3D cell culture system of human A549 lung cell line plus sodium alginate was used to create the scaffolds that were infected with Pb18. Our results showed low cytotoxicity, evidence of increased cell density (indicative of cell proliferation), and the maintenance of cell viability for seven days. The confocal analysis revealed viable yeast within the 3D scaffold, as demonstrated in the solid BHI Agar medium cultivation. Moreover, when ECM proteins were added to the alginate scaffolds, the number of retrieved fungi was significantly higher. Our results highlight that this 3D model may be promising for in vitro studies of host–pathogen interactions.

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Cell proliferation in in vivo‐like three‐dimensional cell culture is regulated by sequestration of ERK1/2 to lipid rafts

Abstract: These results imply that under in vivo-like conditions, cells might achieve reduction of their proliferation level by sequestering activated ERK1/2 to lipid rafts.

Cited by 4 publications

References 49 publications

The characteristics of Ishikawa endometrial cancer cells are modified by substrate topography with cell-like features and the polymer surface

The characteristics of Ishikawa endometrial cancer cells are modified by substrate topography with cell-like features and the polymer surface

Quercetin affects membrane lipids and apoptosis in three-dimensional fibroblast cultures

Alginate-Based 3D A549 Cell Culture Model to Study Paracoccidioides Infection

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