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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

Atomic force microscopy (AFM) manipulation of cultured cells.(A) Chemical structure of the BAM molecule. It comprises an oleyl group, an NHS-reactive ester group, and a hydrophilic PEG linker. (B) Diagrams of AFM manipulation of the adhered or BAM-anchored suspended cells (upper images). Phase-contrast micrographs (lower images) of adherent and BAM-anchored suspended HEK293 cells (the deltoidal object on the left is the AFM cantilever).
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fig-1: Atomic force microscopy (AFM) manipulation of cultured cells.(A) Chemical structure of the BAM molecule. It comprises an oleyl group, an NHS-reactive ester group, and a hydrophilic PEG linker. (B) Diagrams of AFM manipulation of the adhered or BAM-anchored suspended cells (upper images). Phase-contrast micrographs (lower images) of adherent and BAM-anchored suspended HEK293 cells (the deltoidal object on the left is the AFM cantilever).

Mentions: The pyramidal probe (SN-AF01S-NT; spring constant: 0.02 N/m) was purchased from Seiko Instruments Inc. (Tokyo, Japan). Human embryonic kidney HEK293 cells were obtained from Health Science Research Resources Bank (Osaka, Japan). Cell anchoring molecule, BAM (SUNBRIGHT OE-020CS) (Fig. 1A), was purchased from NOF Corporation (Tokyo, Japan). F-actin labeling kit was purchased from AAT Bioquest, Inc. (Sunnyvale, California, USA). Other reagents were purchased from Sigma-Aldrich (St. Louis, Missouri, USA), Wako Pure Chemical Industries Ltd. (Osaka, Japan), or Life Technologies Japan Ltd. (Tokyo, Japan).


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

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

Atomic force microscopy (AFM) manipulation of cultured cells.(A) Chemical structure of the BAM molecule. It comprises an oleyl group, an NHS-reactive ester group, and a hydrophilic PEG linker. (B) Diagrams of AFM manipulation of the adhered or BAM-anchored suspended cells (upper images). Phase-contrast micrographs (lower images) of adherent and BAM-anchored suspended HEK293 cells (the deltoidal object on the left is the AFM cantilever).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig-1: Atomic force microscopy (AFM) manipulation of cultured cells.(A) Chemical structure of the BAM molecule. It comprises an oleyl group, an NHS-reactive ester group, and a hydrophilic PEG linker. (B) Diagrams of AFM manipulation of the adhered or BAM-anchored suspended cells (upper images). Phase-contrast micrographs (lower images) of adherent and BAM-anchored suspended HEK293 cells (the deltoidal object on the left is the AFM cantilever).
Mentions: The pyramidal probe (SN-AF01S-NT; spring constant: 0.02 N/m) was purchased from Seiko Instruments Inc. (Tokyo, Japan). Human embryonic kidney HEK293 cells were obtained from Health Science Research Resources Bank (Osaka, Japan). Cell anchoring molecule, BAM (SUNBRIGHT OE-020CS) (Fig. 1A), was purchased from NOF Corporation (Tokyo, Japan). F-actin labeling kit was purchased from AAT Bioquest, Inc. (Sunnyvale, California, USA). Other reagents were purchased from Sigma-Aldrich (St. Louis, Missouri, USA), Wako Pure Chemical Industries Ltd. (Osaka, Japan), or Life Technologies Japan Ltd. (Tokyo, Japan).

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