Simulated Microgravity Culture System for a 3-D Carcinoma Tissue Model

Nakamura, Katsuya; Kuga, Hirotaka; Morisaki, Takafumi; Baba, Elio H.; Sato, Norihiro; Mizumoto, Kazuhiro; Sueishi, Katsuo; Tanaka, Masato

doi:10.2144/02335rr02

Cited by 31 publications

(18 citation statements)

References 30 publications

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“…Herein we investigate if a similar pattern could be retrieved in breast cancer cells and how such features are associated with differences in their biochemical pathways. Indeed, conflicting data have been reported by investigations carried out on cancer cells exposed to microgravity: some authors have recorded an overall inhibitory effect on cancer cell proliferation, motility, and survival [18, 19], whereas others have observed the opposite [20–22]. We hypothesize that such results may be likely explained by the emergence of distinct cell phenotypes, characterized by different functional and reproductive features.…”

Section: Introductionmentioning

confidence: 68%

Phenotypic Switch Induced by Simulated Microgravity on MDA-MB-231 Breast Cancer Cells

Masiello

Cucina

Proietti

et al. 2014

BioMed Research International

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Microgravity exerts dramatic effects on cell morphology and functions, by disrupting cytoskeleton and adhesion structures, as well as by interfering with biochemical pathways and gene expression. Impairment of cells behavior has both practical and theoretical significance, given that investigations of mechanisms involved in microgravity-mediated effects may shed light on how biophysical constraints cooperate in shaping complex living systems. By exposing breast cancer MDA-MB-231 cells to simulated microgravity (~0.001 g), we observed the emergence of two morphological phenotypes, characterized by distinct membrane fractal values, surface area, and roundness. Moreover, the two phenotypes display different aggregation profiles and adherent behavior on the substrate. These morphological differences are mirrored by the concomitant dramatic functional changes in cell processes (proliferation and apoptosis) and signaling pathways (ERK, AKT, and Survivin). Furthermore, cytoskeleton undergoes a dramatic reorganization, eventually leading to a very different configuration between the two populations. These findings could be considered adaptive and reversible features, given that, by culturing microgravity-exposed cells into a normal gravity field, cells are enabled to recover their original phenotype. Overall these data outline the fundamental role gravity plays in shaping form and function in living systems.

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Section: Introductionmentioning

confidence: 68%

Phenotypic Switch Induced by Simulated Microgravity on MDA-MB-231 Breast Cancer Cells

Masiello

Cucina

Proietti

et al. 2014

BioMed Research International

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“…The cytoskeleton, DNA structure, and microtubules of the human breast cancer cell line MCF-7 changed during space flight due to lower proliferation and blockage of the cell cycle (Vassy et al, 2003). However, there was less apoptosis and increased mitosis observed in human pancreas carcinoma cells under simulated weightlessness using a clinostat (Nakamura et al, 2002). Taga et al (2006) indicated that the simulated microgravity conditions may have altered murine B16-F10 melanoma cell characteristics and enhanced their invasive properties.…”

Section: Cellmentioning

confidence: 99%

Effects of simulated weightlessness on cellular morphology and biological characteristics of cell lines SGC-7901 and HFE-145

Zhu

Jin

et al. 2014

Genet. Mol. Res.

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ABSTRACT. We investigated the effects of simulated weightlessness on cellular morphology, proliferation, cell cycle, and apoptosis of the human gastric carcinoma cell line SGC-7901 and the human gastric normal cell line HFE-145. A rotating clinostat was used to simulate weightlessness. The Image-Pro4.5 image analysis system was used for morphometric analysis. Proliferating cell nuclear antigen expression was examined by immunohistochemical staining. Changes in the cell cycle were examined using a cytometer. Apoptosis was measured using the terminal dUTP nick-end labeling (TUNEL) method. When subjected to simulated weightlessness, the cellular morphology of SGC-7901 cells was changed at 12, 24, 48, and 72 h, cell conversion from the G 1 to S phase was blocked, proliferation was inhibited at 48 and 72 h, and the apoptosis index was increased at 72 h. The same changes were observed for HFE-145 cells at 12 h when subjected to simulated weightlessness, but no significant changes were found 6061 ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 13 (3): 6060-6069 (2014) Effects of simulated weightlessness on cellular morphology afterward compared with controls. SGC-7901 cells change their cellular morphology and biological characteristics during clinostat-simulated weightlessness at 72 h, but HFE-145 cells only change at 12 h and adapt to simulated weightlessness after that point.

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“…In this study, we have examined some morphological and biochemical events on pachytene spermatocytes cultured in rotating-wall vessels that provide a simulated microgravity condition. This novel in vitro culture technology has been used to improve tissue culture conditions: it was demonstrated that fibroblasts or carcinoma tissues cultured under a simulated microgravity condition show a greater number of mitotic cells and a lower number of apoptotic cells than control cells cultured under gravity condition (Ingram et al 1997, Nakamura et al 2002. At the molecular level, it was also shown that the microgravity culture system leads to upregulation of the expression of mitotic genes in proliferating cells (Nakamura et al 2002).…”

Section: Introductionmentioning

confidence: 99%

“…This novel in vitro culture technology has been used to improve tissue culture conditions: it was demonstrated that fibroblasts or carcinoma tissues cultured under a simulated microgravity condition show a greater number of mitotic cells and a lower number of apoptotic cells than control cells cultured under gravity condition (Ingram et al 1997, Nakamura et al 2002. At the molecular level, it was also shown that the microgravity culture system leads to upregulation of the expression of mitotic genes in proliferating cells (Nakamura et al 2002). Our results on mouse spermatocytes have shown that microgravity culture conditions promote spontaneous meiotic progression, as monitored by the significant increase of chromatin condensation and by molecular parameters such as activation of MAPKs, p90Rsk2 kinase and MPF.…”

Section: Introductionmentioning

confidence: 99%

Meiotic progression of isolated mouse spermatocytes under simulated microgravity

et al. 2004

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Progression through the prophase of the first meiotic division can be obtained in culture by treatment of mouse spermatocytes with the serine/threonine phosphatase inhibitor okadaic acid. Chromosome condensation during this G2/M transition involves the activation of the MAPK pathway, which causes the activation of Nek2 and the phosphorylation of the chromatin architectural protein Hmga2. In an effort to set up conditions to allow a spontaneous progression of mouse spermatocytes through meiosis, we have investigated the cell-cycle features of these cells cultured for 24 h with a rotary cell culture system in a humidified atmosphere in a thermostatic incubator to simulate a microgravity environment. Morphological analysis of nuclear squashes indicated a 2-fold increase in late-pachytene spermatocytes with highly condensed chromosomes, and a contemporaneous decrease of mid-pachytene cells with less condensed chromatin. Microgravity induced a 2-fold activation of the cyclinB-cdc2 complex, confirming at the molecular level that cell-cycle progression had occurred. Moreover, using immunokinase assays with specific substrates we have demonstrated that the meiotic progression obtained under microgravity conditions is accompanied by activation of the Erk1/p90Rsk2 pathway. These data indicated that activation of the MAPK pathway correlates with chromatin condensation even under conditions in which meiotic progression occurs spontaneously and is not induced by a drug. We suggest that culture under microgravity conditions might help to release the block that inhibits isolated spermatocytes from progressing through prophase at unit gravity, and to study the physiological events of germ cell differentiation in vitro.

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Simulated Microgravity Culture System for a 3-D Carcinoma Tissue Model

Cited by 31 publications

References 30 publications

Phenotypic Switch Induced by Simulated Microgravity on MDA-MB-231 Breast Cancer Cells

Phenotypic Switch Induced by Simulated Microgravity on MDA-MB-231 Breast Cancer Cells

Effects of simulated weightlessness on cellular morphology and biological characteristics of cell lines SGC-7901 and HFE-145

Meiotic progression of isolated mouse spermatocytes under simulated microgravity

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