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Distinct mechanical behavior of HEK293 cells in adherent and suspended states.

Haghparast SM, Kihara T, Miyake J - PeerJ (2015)

Bottom Line: The mechanical features of individual animal cells have been regarded as indicators of cell type and state.In this paper, we report the unique mechanical and actin cytoskeletal features of human embryonic kidney HEK293 cells.Induced actin filament depolymerization revealed that the actin cytoskeleton was the underlying source of the stiffness in suspended HEK293 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University , Toyonaka, Osaka , Japan.

ABSTRACT
The mechanical features of individual animal cells have been regarded as indicators of cell type and state. Previously, we investigated the surface mechanics of cancer and normal stromal cells in adherent and suspended states using atomic force microscopy. Cancer cells possessed specific mechanical and actin cytoskeleton features that were distinct from normal stromal cells in adherent and suspended states. In this paper, we report the unique mechanical and actin cytoskeletal features of human embryonic kidney HEK293 cells. Unlike normal stromal and cancer cells, the surface stiffness of adherent HEK293 cells was very low, but increased after cell detachment from the culture surface. Induced actin filament depolymerization revealed that the actin cytoskeleton was the underlying source of the stiffness in suspended HEK293 cells. The exclusive mechanical response of HEK293 cells to perturbation of the actin cytoskeleton resembled that of adherent cancer cells and suspended normal stromal cells. Thus, with respect to their special cell-surface mechanical features, HEK293 cells could be categorized into a new class distinct from normal stromal and cancer cells.

No MeSH data available.


Related in: MedlinePlus

Confocal laser-scanning microscopy (CLSM) images of fluorescently labeled F-actin of adhered and BAM-anchored suspended HEK293 cells.Superimposed images of the apical cell surface (Apical), middle part (Middle), and whole cell (Whole) are shown. The thickness of superimposed images (z) is shown individually.
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fig-2: Confocal laser-scanning microscopy (CLSM) images of fluorescently labeled F-actin of adhered and BAM-anchored suspended HEK293 cells.Superimposed images of the apical cell surface (Apical), middle part (Middle), and whole cell (Whole) are shown. The thickness of superimposed images (z) is shown individually.

Mentions: Furthermore, the actin cytoskeleton structures of HEK293 cells cultured under these conditions were observed by CLSM (Fig. 2). Apparently, there was not much difference in F-actin structures of HEK293 cells in the adherent and suspended states. Immature F-actin structures were observed on the apical surface of both spread and spherical cells. The peripheral F-actin structures at the plasma membrane were observed in the middle part of the adherent cells. In suspended cells, a clear ring-shaped cortical actin was visible in the middle part of the serial image. Whole-cell imaging revealed numerous dot-shaped F-actin structures inside the adherent cells, while a lot of projections appeared on the surface of suspended cells. No developed actin stress fibers were noticed in both adhesion states. Thus, with respect to the possible observations of the F-actin structures in HEK293 cells by CLSM, the structures appeared to be immature by nature and seemed to be unchanged after removing the cells from the culture surface.


Distinct mechanical behavior of HEK293 cells in adherent and suspended states.

Haghparast SM, Kihara T, Miyake J - PeerJ (2015)

Confocal laser-scanning microscopy (CLSM) images of fluorescently labeled F-actin of adhered and BAM-anchored suspended HEK293 cells.Superimposed images of the apical cell surface (Apical), middle part (Middle), and whole cell (Whole) are shown. The thickness of superimposed images (z) is shown individually.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4525692&req=5

fig-2: Confocal laser-scanning microscopy (CLSM) images of fluorescently labeled F-actin of adhered and BAM-anchored suspended HEK293 cells.Superimposed images of the apical cell surface (Apical), middle part (Middle), and whole cell (Whole) are shown. The thickness of superimposed images (z) is shown individually.
Mentions: Furthermore, the actin cytoskeleton structures of HEK293 cells cultured under these conditions were observed by CLSM (Fig. 2). Apparently, there was not much difference in F-actin structures of HEK293 cells in the adherent and suspended states. Immature F-actin structures were observed on the apical surface of both spread and spherical cells. The peripheral F-actin structures at the plasma membrane were observed in the middle part of the adherent cells. In suspended cells, a clear ring-shaped cortical actin was visible in the middle part of the serial image. Whole-cell imaging revealed numerous dot-shaped F-actin structures inside the adherent cells, while a lot of projections appeared on the surface of suspended cells. No developed actin stress fibers were noticed in both adhesion states. Thus, with respect to the possible observations of the F-actin structures in HEK293 cells by CLSM, the structures appeared to be immature by nature and seemed to be unchanged after removing the cells from the culture surface.

Bottom Line: The mechanical features of individual animal cells have been regarded as indicators of cell type and state.In this paper, we report the unique mechanical and actin cytoskeletal features of human embryonic kidney HEK293 cells.Induced actin filament depolymerization revealed that the actin cytoskeleton was the underlying source of the stiffness in suspended HEK293 cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University , Toyonaka, Osaka , Japan.

ABSTRACT
The mechanical features of individual animal cells have been regarded as indicators of cell type and state. Previously, we investigated the surface mechanics of cancer and normal stromal cells in adherent and suspended states using atomic force microscopy. Cancer cells possessed specific mechanical and actin cytoskeleton features that were distinct from normal stromal cells in adherent and suspended states. In this paper, we report the unique mechanical and actin cytoskeletal features of human embryonic kidney HEK293 cells. Unlike normal stromal and cancer cells, the surface stiffness of adherent HEK293 cells was very low, but increased after cell detachment from the culture surface. Induced actin filament depolymerization revealed that the actin cytoskeleton was the underlying source of the stiffness in suspended HEK293 cells. The exclusive mechanical response of HEK293 cells to perturbation of the actin cytoskeleton resembled that of adherent cancer cells and suspended normal stromal cells. Thus, with respect to their special cell-surface mechanical features, HEK293 cells could be categorized into a new class distinct from normal stromal and cancer cells.

No MeSH data available.


Related in: MedlinePlus