Altered Distribution of Mitochondria and Actin Fibers in 3T3 Cells Cultured on Microcarriers

Laney, Tracey V. du; Cherry, Robert; Coppinger, Justine A.; Truskey, George A.

doi:10.1021/bp00018a600

Cited by 5 publications

(4 citation statements)

References 18 publications

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“…The relationship between the organization of actin cytoskeleton and distribution of mitochondria attributes to normal cell physiology. 49 We observed that mitochondria distribution was parallel to the direction of actin fibers. Overall, this study demonstrates the suitability of the model to retain cell functionality after the freeze−thaw cycle.…”

Section: Resultsmentioning

confidence: 69%

“…Interestingly, MitoTracker Red CMXRos staining did not show significant differences in mitochondria distribution between a fresh and frozen/thawed cell (Figure B,E). The relationship between the organization of actin cytoskeleton and distribution of mitochondria attributes to normal cell physiology . We observed that mitochondria distribution was parallel to the direction of actin fibers.…”

Section: Resultsmentioning

confidence: 75%

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Cryoprotectant-Free Freezing of Cells Using Liquid Marbles Filled with Hydrogel

Vadivelu

Kashaninejad

Sreejith

et al. 2018

ACS Appl. Mater. Interfaces

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Cryopreservation without cryoprotectant remains a significant challenge for the re-establishment of cell culture after freeze−thaw. Thus, finding an alternative and a simple cryopreservation method is necessary. Liquid marble (LM)-based digital microfluidics is a promising approach for cryoprotectant-free cryopreservation. However, the use of this platform to efficiently preserve samples with low cell density and well-controlled serum concentrations has not been investigated. We addressed this issue by embedding an agarose-containing fetal bovine serum (FBS) inside the LM. A low density of 500 cells/μL of murine 3T3 cells was selected for evaluating the postcryogenic survivability. The effects on the post-thaw cell viability of the concentration of agarose, the amount of FBS inside the agarose, and the volume of the LM were investigated systematically. This paper also presents an analysis on the changes in shape and crack size of post-thawed agarose. The results revealed that the embedded agarose gel serves as a controlled release mechanism of FBS and significantly improves cell viability. Post-thaw recovery sustains major cellular features, such as viability, cell adhesion, and morphology. The platform technology reported here opens up new possibilities to cryopreserve rare biological samples without the toxicity risk of cryoprotectants.

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Section: Resultsmentioning

confidence: 69%

Section: Resultsmentioning

confidence: 75%

Cryoprotectant-Free Freezing of Cells Using Liquid Marbles Filled with Hydrogel

Vadivelu

Kashaninejad

Sreejith

et al. 2018

ACS Appl. Mater. Interfaces

View full text Add to dashboard Cite

show abstract

“…According to this rule, poor absorption or permeation is more likely when there are more than five hydrogen bond donors, ten hydrogen bond acceptors, the molecular weight is greater than 500 g/mol and the calculated log p (logarithmic ratio of the octanol–water partitioning coefficient) is greater than 5. Molecules violating more than one of these parameters may have problems with bioavailability and a high probability of failure to display drug-likeness [ 15 ]. Further, the topological polar surface area (TPSA), which is another key property in estimating drug bioavailability, was also calculated.…”

Section: Resultsmentioning

confidence: 99%

“…This model is described in the literature and is widely used to study the resistance of mammalian cells to bacteria and the action of various chemicals, including antioxidants, in normal and pathological conditions [ 13 ]. Moreover, these cell lines are widely used to study the structure of the cytoskeleton [ 14 , 15 ]. It was shown that under the action of N-acetylcysteine, there is a change in the actin cytoskeleton structure in 3T3-SV40 typical to normal cells [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

Identification of Spiro-Fused [3-azabicyclo[3.1.0]hexane]oxindoles as Potential Antitumor Agents: Initial In Vitro Evaluation of Anti-Proliferative Effect and Actin Cytoskeleton Transformation in 3T3 and 3T3-SV40 Fibroblast

Knyazev

Shmakov

Pechkovskaya

et al. 2021

IJMS

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Novel heterocyclic compounds containing 3-spiro[3-azabicyclo[3.1.0]hexane]oxindole framework (4a, 4b and 4c) have been studied as potential antitumor agents. The in silico ADMET (adsorption, distribution, metabolism, excretion and toxicity) analysis was performed on 4a–c compounds with promising antiproliferative activity, previously synthetized and screened against human erythroleukemic cell line K562 tumor cell line. Cytotoxicity of 4a–c against murine fibroblast 3T3 and SV-40 transformed murine fibroblast 3T3-SV40 cell lines were evaluated. The 4a and 4c compounds were cytotoxic against 3T3-SV40 cells in comparison with those of 3T3. In agreement with the DNA cytometry studies, the tested compounds have achieved significant cell-cycle perturbation with higher accumulation of cells in G0/G1 phase. Using confocal microscopy, we found that with 4a and 4c treatment of 3T3 cells, actin filaments disappeared, and granular actin was distributed diffusely in the cytoplasm in 82–97% of cells. The number of 3T3-SV40 cells with stress fibers increased to 7–30% against 2% in control. We discovered that transformed 3T3-SV40 cells after treatment with compounds 4a and 4c significantly reduced the number of cells with filopodium-like membrane protrusions (from 86 % in control cells to 6–18% after treatment), which indirectly suggests a decrease in cell motility. We can conclude that the studied compounds 4a and 4c have a cytostatic effect, which can lead to a decrease in the number of filopodium-like membrane protrusions.

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A numerical analysis of forces exerted by laminar flow on spreading cells in a parallel plate flow chamber assay

Olivier

Truskey

1993

Biotech & Bioengineering

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Exposure of spreading anchorage-dependent cells to laminar flow is a common technique to measure the strength of cell adhesion. Since cells protrude into the flow stream, the force exerted by the fluid on the cells is a function of cell shape. To assess the relationship between cell shape and the hydrodynamic force on adherent cells, we obtained numerical solutions of the velocity and stress fields around bovine aortic endothelial cells during various stages of spreading and calculated the force required to detach the cells. Morphometric parameters were obtained from light and scanning electron microscopy measurements. Cells were assumed to have a constant volume, but the surface area increased during spreading until the membrane was stretched taut. Two-dimensional models of steady flow were generated using the software packages ANSYS (mesh generation) and FIDAP (problem solution). The validity of the numerical results was tested by comparison with published results for a semicircle in contact with the surface. The drag force and torque were greatest for round cells making initial contact with the surface. During spreading, the drag force and torque declined by factors of 2 and 20, respectively. The calculated forces and moments were used in adhesion models to predict the wall shear stress at which the cells detached. Based upon published values for the bond force and receptor number, round cells should detach at shear stresses between 2.5 and 6 dyn/cm(2), whereas substantially higher stresses are needed to detach spreading and fully spread cells. Results from the simulations indicate that (1) the drag force varies little with cell shape whereas the torque is very sensitive to cell shape, and (2) the increase in the strength of adhesion during spreading is due to increased contact area and receptor densities within the contact area.

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Altered Distribution of Mitochondria and Actin Fibers in 3T3 Cells Cultured on Microcarriers

Cited by 5 publications

References 18 publications

Cryoprotectant-Free Freezing of Cells Using Liquid Marbles Filled with Hydrogel

Cryoprotectant-Free Freezing of Cells Using Liquid Marbles Filled with Hydrogel

Identification of Spiro-Fused [3-azabicyclo[3.1.0]hexane]oxindoles as Potential Antitumor Agents: Initial In Vitro Evaluation of Anti-Proliferative Effect and Actin Cytoskeleton Transformation in 3T3 and 3T3-SV40 Fibroblast

A numerical analysis of forces exerted by laminar flow on spreading cells in a parallel plate flow chamber assay

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