Relationship between macrophage differentiation and the chemotactic activity toward damaged myoblast cells

Uchida, Masataka; Oyanagi, Eri; Miyachi, Motohiko; Yamauchi, Akira; Yano, Hiromi

doi:10.1016/j.jim.2013.04.007

Cited by 4 publications

(10 citation statements)

References 27 publications

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“…TAXIScan is an assay device for studying cell dynamics in vitro, which functions by providing two-dimensional images of cell migration. Accordingly, this device can provide markedly more information, including morphology as well as quantitative analysis, compared to existing methods such as the Boyden chamber method [ 13 , 24 , 25 , 26 ]. Interestingly, the directionality and velocity of chemotactic cells can be provided by analyzing the cell images.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, the velocity of J774 cells was not different between LPS and LPS+Ly cells. A previous study reported that the chemotactic activity of M1 macrophages to damaged skeletal muscle cells is lower than that of M2 macrophages, which is more directionally dependent than velocity [ 13 ]. Expressions of Cd11c and Tlr4 mRNA as markers of pro-inflammatory M1 macrophages have been demonstrated [ 24 , 27 ].…”

Section: Discussionmentioning

confidence: 99%

“…The C2C12 cells were used from passage 2 to 10. The killed-C2C12 cells were induced by repeat freezing (three times) by liquid nitrogen for 60 sec and incubation at 37 °C for 3 min, and then they were combined with live C2C12 at 1:0.5 (live:killed C2C12) as a partially damaged muscle model [ 13 ].…”

Section: Methodsmentioning

confidence: 99%

“…Chemotaxis assays of J774 cells were carried out using the TAXIScan device (ECI, Tokyo, Japan) [ 13 , 23 , 24 ]. Migration of J774 cells at 37 °C was recorded every 3 min.…”

Section: Methodsmentioning

confidence: 99%

“…Kleveta et al (2012) [ 12 ] showed that F-actin expression was upregulated by LPS treatment, and moreover, we reported that LPS-treated macrophages were accompanied by both a high F-actin expression with a low expression of CCR2 (a receptor for monocyte chemoattractant protein-1) and a decrease in chemotactic activity [ 13 ]. Therefore, macrophage differentiation strongly influenced the chemotactic activity toward damaged myoblast cells through the expression of F-actin [ 13 ] and CCR2 [ 13 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

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Reduction of Real-Time Imaging of M1 Macrophage Chemotaxis toward Damaged Muscle Cells is PI3K-Dependent

et al. 2018

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Macrophages migrate and invade into damaged muscle rapidly and are important for muscle repair and subsequent regeneration. The exact cellular and biological events that cause macrophage migration toward injured muscle are not completely understood. In this study, the effect of macrophage differentiation on the chemotactic capability to invade local damaged muscle was investigated using an in vitro model of muscle injury. We used C2C12 cell myoblasts and J774 cell macrophages, and the “killed-C2C12” cells were combined with live C2C12 cells as a partially damaged muscle model. The cultured J774 cells, with or without lipopolysaccharide (LPS), were treated with Ly294002 (Ly), which is an inhibitor of phosphoinositide 3-kinase (PI3K). In order to evaluate the polarization effect of LPS stimulation on J774 cells, expression of cell surface Toll-like receptor 4 (TLR4), CD11c and CCR2, and expression of F-actin intensity, were analyzed by flow cytometry. The real-time horizontal chemotaxis assay of J774 cells was tested using the TAXIScan device. The expressions of TLR4, CD11c, and F-actin intensity in LPS-treated cells were significantly higher than those in Ctrl cells. In LPS-treated cells, the chemotactic activity toward damaged muscle cells completely disappeared. Moreover, the reduced chemotaxis depended far more on directionality than velocity. However, Ly treatment reversed the reduced chemotactic activity of the LPS-treated cells. In addition, cell-adhesion and F-actin intensity, but not CCR2 expression, in LPS-treated cells, was significantly reduced by Ly treatment. Taken together, our results suggest that the PI3K/Akt activation state drives migration behavior towards damaged muscle cells.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…Chemotaxis assays of J774 cells were carried out using the TAXIScan device (ECI, Tokyo, Japan) [ 13 , 23 , 24 ]. Migration of J774 cells at 37 °C was recorded every 3 min.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reduction of Real-Time Imaging of M1 Macrophage Chemotaxis toward Damaged Muscle Cells is PI3K-Dependent

et al. 2018

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STIM1-dependent store-operated calcium entry mediates sex differences in macrophage chemotaxis and monocyte recruitment

Fresquez,

Hogan,

Rivera

et al. 2024

Journal of Biological Chemistry

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Evaluation of pancreatic cancer cell migration with multiple parameters in vitro by using an optical real-time cell mobility assay device

et al. 2017

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BackgroundMigration of cancer cell correlates with distant metastasis and local invasion, which are good targets for cancer treatment. An optically accessible device “TAXIScan” was developed, which provides considerably more information regarding the cellular dynamics and less quantity of samples than do the existing methods. Here, we report the establishment of a system to analyze the nature of pancreatic cancer cells using TAXIScan and we evaluated lysophosphatidic acid (LPA)-elicited pancreatic cell migration.MethodsPancreatic cancer cell lines, BxPC3, PANC-1, AsPC1, and MIAPaCa-2, were analyzed for adhesion as well as migration towards LPA by TAXIScan using parameters such as velocity and directionality or for the number of migrated cells by the Boyden chamber methods. To confirm that the migration was initiated by LPA, the expression of LPA receptors and activation of intracellular signal transductions were examined by quantitative reverse transcriptase polymerase reaction and western blotting.ResultsScaffold coating was necessary for the adhesion of pancreatic cancer cells, and collagen I and Matrigel were found to be good scaffolds. BxPC3 and PANC-1 cells clearly migrated towards the concentration gradient formed by injecting 1 μL LPA, which was abrogated by pre-treatment with LPA inhibitor, Ki16425 (IC50 for the directionality ≈ 1.86 μM). The LPA dependent migration was further confirmed by mRNA and protein expression of LPA receptors as well as phosphorylation of signaling molecules. LPA1 mRNA was highest among the 6 receptors, and LPA1, LPA2 and LPA3 proteins were detected in BxPC3 and PANC-1 cells. Phosphorylation of Akt (Thr308 and Ser473) and p42/44MAPK in BxPC3 and PANC-1 cells was observed after LPA stimulation, which was clearly inhibited by pre-treatment with a compound Ki16425.ConclusionsWe established a novel pancreatic cancer cell migration assay system using TAXIScan. This assay device provides multiple information on migrating cells simultaneously, such as their morphology, directionality, and velocity, with a small volume of sample and can be a powerful tool for analyzing the nature of cancer cells and for identifying new factors that affect cell functions.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-017-3218-4) contains supplementary material, which is available to authorized users.

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Relationship between macrophage differentiation and the chemotactic activity toward damaged myoblast cells

Cited by 4 publications

References 27 publications

Reduction of Real-Time Imaging of M1 Macrophage Chemotaxis toward Damaged Muscle Cells is PI3K-Dependent

Reduction of Real-Time Imaging of M1 Macrophage Chemotaxis toward Damaged Muscle Cells is PI3K-Dependent

STIM1-dependent store-operated calcium entry mediates sex differences in macrophage chemotaxis and monocyte recruitment

Evaluation of pancreatic cancer cell migration with multiple parameters in vitro by using an optical real-time cell mobility assay device

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