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Mitotic cells contract actomyosin cortex and generate pressure to round against or escape epithelial confinement.

Sorce B, Escobedo C, Toyoda Y, Stewart MP, Cattin CJ, Newton R, Banerjee I, Stettler A, Roska B, Eaton S, Hyman AA, Hierlemann A, Müller DJ - Nat Commun (2015)

Bottom Line: Cells that cannot round against nor escape confinement cannot orient their mitotic spindles and more likely undergo apoptosis.The results highlight how spatially constrained epithelial cells prepare for mitosis: either they are strong enough to round up or they must escape.The ability to escape from confinement and reintegrate after mitosis appears to be a basic property of epithelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zurich, Mattenstrasse 26, Basel 4058, Switzerland.

ABSTRACT
Little is known about how mitotic cells round against epithelial confinement. Here, we engineer micropillar arrays that subject cells to lateral mechanical confinement similar to that experienced in epithelia. If generating sufficient force to deform the pillars, rounding epithelial (MDCK) cells can create space to divide. However, if mitotic cells cannot create sufficient space, their rounding force, which is generated by actomyosin contraction and hydrostatic pressure, pushes the cell out of confinement. After conducting mitosis in an unperturbed manner, both daughter cells return to the confinement of the pillars. Cells that cannot round against nor escape confinement cannot orient their mitotic spindles and more likely undergo apoptosis. The results highlight how spatially constrained epithelial cells prepare for mitosis: either they are strong enough to round up or they must escape. The ability to escape from confinement and reintegrate after mitosis appears to be a basic property of epithelial cells.

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Viability and apoptosis of MDCK cells confined by micropillars.(a) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of MDCK cells cultured on micropillar arrays for 3 h in the presence of different chemical compounds that perturb cellular processes. For each condition, mean and s.d. are given. (b) Percentage of apoptotic (red) and living (green) cells on the flat surface in the presence of different perturbants (ncells=120, 140, 80, 95, 100, 113, 105, 95, 80). (c) Percentage of apoptotic and living cells on micropillar arrays (ncells=200, 150, 180, 210, 220, 150, 170, 210, 170). Apoptotic and living cells were quantified using CellProfiler (see Methods). Micropillar arrays used had average interpillar distances of ≈6.8 μm. Mann–Whitney P values indicate the significance relative to the control (untreated) cells. *P<0.01, **P<0.001. Perturbants are described in Supplementary Table 1.
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f5: Viability and apoptosis of MDCK cells confined by micropillars.(a) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of MDCK cells cultured on micropillar arrays for 3 h in the presence of different chemical compounds that perturb cellular processes. For each condition, mean and s.d. are given. (b) Percentage of apoptotic (red) and living (green) cells on the flat surface in the presence of different perturbants (ncells=120, 140, 80, 95, 100, 113, 105, 95, 80). (c) Percentage of apoptotic and living cells on micropillar arrays (ncells=200, 150, 180, 210, 220, 150, 170, 210, 170). Apoptotic and living cells were quantified using CellProfiler (see Methods). Micropillar arrays used had average interpillar distances of ≈6.8 μm. Mann–Whitney P values indicate the significance relative to the control (untreated) cells. *P<0.01, **P<0.001. Perturbants are described in Supplementary Table 1.

Mentions: We have observed that mitotic cells generate an outward-directed force to round up against mechanical confinement. If mechanical confinement is too great, mitotic cells employ this rounding force to escape the confinement. We next sought to study the consequences in cases where mitotic cells were unable to escape the confinement of the micropillars. Therefore, we perturbed the actomyosin cortex or the hydrostatic pressure of MDCK cells seeded on cell culture plates or on micropillar arrays with interpillar distance of ≈6.8 μm and characterized cell viability and apoptosis rates (Fig. 5). The direct comparison of MDCK cells being perturbed in the absence and presence of pillars, showed that MDCK cells that cannot escape too narrow micropillar confinements are more likely to go through apoptosis.


Mitotic cells contract actomyosin cortex and generate pressure to round against or escape epithelial confinement.

Sorce B, Escobedo C, Toyoda Y, Stewart MP, Cattin CJ, Newton R, Banerjee I, Stettler A, Roska B, Eaton S, Hyman AA, Hierlemann A, Müller DJ - Nat Commun (2015)

Viability and apoptosis of MDCK cells confined by micropillars.(a) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of MDCK cells cultured on micropillar arrays for 3 h in the presence of different chemical compounds that perturb cellular processes. For each condition, mean and s.d. are given. (b) Percentage of apoptotic (red) and living (green) cells on the flat surface in the presence of different perturbants (ncells=120, 140, 80, 95, 100, 113, 105, 95, 80). (c) Percentage of apoptotic and living cells on micropillar arrays (ncells=200, 150, 180, 210, 220, 150, 170, 210, 170). Apoptotic and living cells were quantified using CellProfiler (see Methods). Micropillar arrays used had average interpillar distances of ≈6.8 μm. Mann–Whitney P values indicate the significance relative to the control (untreated) cells. *P<0.01, **P<0.001. Perturbants are described in Supplementary Table 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4696517&req=5

f5: Viability and apoptosis of MDCK cells confined by micropillars.(a) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay of MDCK cells cultured on micropillar arrays for 3 h in the presence of different chemical compounds that perturb cellular processes. For each condition, mean and s.d. are given. (b) Percentage of apoptotic (red) and living (green) cells on the flat surface in the presence of different perturbants (ncells=120, 140, 80, 95, 100, 113, 105, 95, 80). (c) Percentage of apoptotic and living cells on micropillar arrays (ncells=200, 150, 180, 210, 220, 150, 170, 210, 170). Apoptotic and living cells were quantified using CellProfiler (see Methods). Micropillar arrays used had average interpillar distances of ≈6.8 μm. Mann–Whitney P values indicate the significance relative to the control (untreated) cells. *P<0.01, **P<0.001. Perturbants are described in Supplementary Table 1.
Mentions: We have observed that mitotic cells generate an outward-directed force to round up against mechanical confinement. If mechanical confinement is too great, mitotic cells employ this rounding force to escape the confinement. We next sought to study the consequences in cases where mitotic cells were unable to escape the confinement of the micropillars. Therefore, we perturbed the actomyosin cortex or the hydrostatic pressure of MDCK cells seeded on cell culture plates or on micropillar arrays with interpillar distance of ≈6.8 μm and characterized cell viability and apoptosis rates (Fig. 5). The direct comparison of MDCK cells being perturbed in the absence and presence of pillars, showed that MDCK cells that cannot escape too narrow micropillar confinements are more likely to go through apoptosis.

Bottom Line: Cells that cannot round against nor escape confinement cannot orient their mitotic spindles and more likely undergo apoptosis.The results highlight how spatially constrained epithelial cells prepare for mitosis: either they are strong enough to round up or they must escape.The ability to escape from confinement and reintegrate after mitosis appears to be a basic property of epithelial cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zurich, Mattenstrasse 26, Basel 4058, Switzerland.

ABSTRACT
Little is known about how mitotic cells round against epithelial confinement. Here, we engineer micropillar arrays that subject cells to lateral mechanical confinement similar to that experienced in epithelia. If generating sufficient force to deform the pillars, rounding epithelial (MDCK) cells can create space to divide. However, if mitotic cells cannot create sufficient space, their rounding force, which is generated by actomyosin contraction and hydrostatic pressure, pushes the cell out of confinement. After conducting mitosis in an unperturbed manner, both daughter cells return to the confinement of the pillars. Cells that cannot round against nor escape confinement cannot orient their mitotic spindles and more likely undergo apoptosis. The results highlight how spatially constrained epithelial cells prepare for mitosis: either they are strong enough to round up or they must escape. The ability to escape from confinement and reintegrate after mitosis appears to be a basic property of epithelial cells.

Show MeSH
Related in: MedlinePlus