A ring barrier–based migration assay to assess cell migration in vitro

2015 ◽  
Vol 10 (6) ◽  
pp. 904-915 ◽  
Author(s):  
Asha M Das ◽  
Alexander M M Eggermont ◽  
Timo L M ten Hagen
Keyword(s):  
Cell Migration ◽  
Migration Assay

scholarly journals APC binds intermediate filaments and is required for their reorganization during cell migration

2013 ◽  
Vol 200 (3) ◽  
pp. 249-258 ◽  
Author(s):  
Yasuhisa Sakamoto ◽  
Batiste Boëda ◽  
Sandrine Etienne-Manneville
Keyword(s):  
Cell Migration ◽  
Glial Fibrillary Acidic Protein ◽  
Intermediate Filaments ◽  
Carcinoma Cells ◽  
Adenomatous Polyposis ◽  
Polyposis Coli ◽  
Cellular Functions ◽  
Migration Assay ◽  
Armadillo Repeats

Intermediate filaments (IFs) are components of the cytoskeleton involved in most cellular functions, including cell migration. Primary astrocytes mainly express glial fibrillary acidic protein, vimentin, and nestin, which are essential for migration. In a wound-induced migration assay, IFs reorganized to form a polarized network that was coextensive with microtubules in cell protrusions. We found that the tumor suppressor adenomatous polyposis coli (APC) was required for microtubule interaction with IFs and for microtubule-dependent rearrangements of IFs during astrocyte migration. We also show that loss or truncation of APC correlated with the disorganization of the IF network in glioma and carcinoma cells. In migrating astrocytes, vimentin-associated APC colocalized with microtubules. APC directly bound polymerized vimentin via its armadillo repeats. This binding domain promoted vimentin polymerization in vitro and contributed to the elongation of IFs along microtubules. These results point to APC as a crucial regulator of IF organization and confirm its fundamental role in the coordinated regulation of cytoskeletons.

119 FGF2 AND FGF10 STIMULATES BOVINE AND OVINE TROPHOBLAST CELL MIGRATION

2011 ◽  
Vol 23 (1) ◽  
pp. 164
Author(s):  
Q. E. Yang ◽  
K. Zhang ◽  
M. I. Giassetti ◽  
M. Ozawa ◽  
S. E. Johnson ◽  
...  
Keyword(s):  
Cell Migration ◽  
Growth Factors ◽  
P38 Mapk ◽  
Trophoblast Cell ◽  
Mitogen Activated Protein Kinase ◽  
Epifluorescence Microscopy ◽  
Migration And Invasion ◽  
Serum Starvation ◽  
Migration Assay

Following hatching, bovine and ovine conceptuses undergo a phase of massive development and remodelling that causes elongation and filamentation. Proper trophoblast cell development and interaction with the uterus is critical for the establishment and maintenance of pregnancy. Various growth factors, including several fibroblast growth factors (FGFs), are produced by the uterus, and at least two of these, FGF2 and FGF10, are released into uterus lumen during early pregnancy. Microarray analysis found that gene products associated with migration and invasion were altered in bovine blastocysts exposed to FGF2 or 10. The objective of this work was to determine if FGF2 and FGF10 impact bovine and ovine trophoblast cell migration. The ability of FGF2 and FGF10 to influence migratory ability of trophoblast cells was examined by using an in vitro transwell migration assay. The bovine trophoblast line, CT1, was used in the first study. After serum starvation, CT1 cells were seeded on the top of each transwell membrane (50 000/transwell) in the presence of vehicle, 0.5, 5, or 50 ng mL–1 bovine recombinant FGF2 or human recombinant FGF10. After 12 h, the transwell was fixed and stained with Hoechst 33342 (0.5 μg mL–1). Migrated cells were counted on five non-overlapping areas of each filter using epifluorescence microscopy. Supplementation with 0.5 ng mL–1 FGF2 increased the number of migrated CT1 cells when compared with controls (268.3 ± 58.3 v. 167.3 ± 47.7; P < 0.01). Supplementation with 5 or 50 ng mL–1 FGF2 further increased the number of migrated CT1 cells (297.0 ± 51.4 and 429.4 ± 98.3, respectively; P < 0.001). Adding 0.5 ng mL–1 FGF10 did not affect CT1 migration but providing 5 or 50 ng mL–1 FGF10 increased CT1 migration (399.8 ± 29.7 and 392.7 ± 58.6 v. 194.2 ± 40.3 for controls; P < 0.005). A subsequent study utilised the ovine trophoblast line, oTR1 in the migration assay (30 000 cells/transwell; 8 h migration assay). Adding 0.5 ng mL–1 FGF2 or FGF10 did not affect oTR1 migration number but exposure to holdout 5 or 50 ng mL–1 FGF2 or FGF10 increased oTR1 migrated cell numbers v. controls (P < 0.05). In a subsequent study, p38 mitogen-activated protein kinase (MAPK), ERK1/2 and JNK signalling cascades utilised by FGF2 and FGF10 in oTR1 cells were investigated. Western blot analysis indicated that both FGF2 and FGF10 induced ERK1/2 and p38 MAPK phosphorylation status. Interestingly FGF10 activated JNK but not p38 MAPK. Taken together, FGF2 and FGF10 stimulate trophoblast cell migration. This response could be mediated by an ERK1/2- or p38 MAPK-dependent system. This project was supported by NRICGP number 2008-35203-19106 from the USDA-NIFA.

scholarly journals Quantification of cancer cell migration with an integrated experimental-computational pipeline

2017 ◽  
Author(s):  
Edwin F. Juarez ◽  
Carolina Garri ◽  
Ahmadreza Ghaffarizadeh ◽  
Paul Macklin ◽  
Kian Kani
Keyword(s):  
Breast Cancer ◽  
Cell Migration ◽  
Cancer Cell ◽  
Cancer Cell Line ◽  
Cancer Cell Migration ◽  
Computational Pipeline ◽  
Migration Assay ◽  
User Friendly ◽  
Migration Of Cells

AbstractWe describe an integrated experimental-computational pipeline for quantifying cell migration in vitro. This pipeline is robust to image noise, open source, and user friendly. The experimental component uses the Oris cell migration assay (Platypus Technologies) to create migration regions. The computational component of the pipeline creates masks in Matlab (MathWorks) to cell-covered regions, uses a genetic algorithm to automatically select the migration region, and outputs a metric to quantify the migration of cells. In this work we demonstrate the utility of our pipeline by quantifying the effects of a drug (Taxol) and of the secreted Anterior Gradient 2 (sAGR2) protein in the migration of MDA-MB-231 cells (a breast cancer cell line). In particular, we show that blocking sAGR2 reduces migration of MDA-MB-231 cells.

A PDMS Device Coupled with Culture Dish for In Vitro Cell Migration Assay

2018 ◽  
Vol 186 (3) ◽  
pp. 633-643 ◽  
Author(s):  
Xiaoqing Lv ◽  
Zhaoxin Geng ◽  
Zhiyuan Fan ◽  
Shicai Wang ◽  
WeiHua Pei ◽  
...  
Keyword(s):  
Cell Migration ◽  
Culture Dish ◽  
Cell Migration Assay ◽  
Migration Assay ◽  
Pdms Device

scholarly journals Adhesion modulates cell morphology and migration within dense fibrous networks

2019 ◽  
Author(s):  
Maurício Moreira-Soares ◽  
Susana P. Cunha ◽  
José Rafael Bordin ◽  
Rui D. M. Travasso
Keyword(s):  
Cell Migration ◽  
Cell Morphology ◽  
Dissipative Particle Dynamics ◽  
Phase Field Model ◽  
Life Forms ◽  
Systematic Analysis ◽  
Regulatory Pathways ◽  
Migration Assay ◽  
Invasion Stage

AbstractOne of the most fundamental abilities required for the sustainability of complex life forms is active cell migration, since it is essential in diverse processes from morphogenesis to leukocyte chemotaxis in immune response. The movement of a cell is the result of intricate mechanisms, that involve the coordination between mechanical forces, biochemical regulatory pathways and environmental cues. In particular, epithelial cancer cells have to employ mechanical strategies in order to migrate through the tissue’s basement membrane and infiltrate the bloodstream during the invasion stage of metastasis. In this work we explore how mechanical interactions such as spatial restriction and adhesion affect migration of a self-propelled droplet in dense fibrous media. We have performed a systematic analysis using a phase-field model and we propose a novel approach to simulate cell migration with Dissipative Particle Dynamics (DPD) modelling. With this purpose we have measured the cell’s velocity and quantified its morphology as a function of the fibre density and of its adhesiveness to the matrix fibres. Furthermore, we have compared our results to a previous in vitro migration assay of fibrosacorma cells in fibrous matrices. The results are model independent and show good agreement between the two methodologies and experiments in the literature, which indicates that these minimalist descriptions are able to capture the main features of the system. Our results indicate that adhesiveness is critical for cell migration, by modulating cell morphology in crowded environments and by enhancing cell velocity. In addition, our analysis suggests that matrix metalloproteinases (MMPs) play an important role as adhesiveness modulators. We propose that new assays should be carried out to address the role of adhesion and the effect of different MMPs in cell migration under confined conditions.

scholarly journals Evaluation of a high-throughput in vitro endothelial cell migration assay for the assessment of nicotine and tobacco delivery products

2020 ◽  
Vol 334 ◽  
pp. 110-116
Author(s):  
Emma Bishop ◽  
Damien Breheny ◽  
Katherine Hewitt ◽  
Mark Taylor ◽  
Tomasz Jaunky ◽  
...  
Keyword(s):  
Cell Migration ◽  
Endothelial Cell ◽  
High Throughput ◽  
Endothelial Cell Migration ◽  
Cell Migration Assay ◽  
Migration Assay

scholarly journals Bacterial Nanocellulose as a Scaffold for In Vitro Cell Migration Assay

Nanomaterials ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2322
Author(s):  
Milena Ugrin ◽  
Jelena Dinic ◽  
Sanja Jeremic ◽  
Sandra Dragicevic ◽  
Bojana Banovic Djeri ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Migration ◽  
Drug Development ◽  
Low Cost ◽  
Boyden Chamber ◽  
Bacterial Nanocellulose ◽  
Cell Migration Assay ◽  
Migration Assay ◽  
Sw480 Cells

Bacterial nanocellulose (BNC) stands out among polymers as a promising biomaterial due to its mechanical strength, hydrophilicity, biocompatibility, biodegradability, low toxicity and renewability. The use of scaffolds based on BNC for 3D cell culture has been previously demonstrated. The study exploited excellent properties of the BNC to develop an efficient and low-cost in vitro cell migration assay. The BNC scaffold was introduced into a cell culture 24 h after the SW480 cells were seeded, and cells were allowed to enter the scaffold within the next 24–48 h. The cells were stained with different fluorophores either before or after the introduction of the scaffold in the culture. Untreated cells were observed to enter the BNC scaffold in significant numbers, form clusters and retain a high viability after 48 h. To validate the assay’s usability for drug development, the treatments of SW480 cells were performed using aspirin, an agent known to reduce the migratory potential of this cell line in culture. This study demonstrates the application of BNC as a scaffold for cell migration testing as a low-cost alternative to commercial assays based on the Boyden chamber principle. The assay could be further developed for routine use in cancer research and anticancer drug development.

scholarly journals A Simplified Method for Quantifying Cell Migration/Wound Healing in 96-Well Plates

2010 ◽  
Vol 15 (4) ◽  
pp. 427-433 ◽  
Author(s):  
Patrick Y. K. Yue ◽  
Emily P. Y. Leung ◽  
N. K. Mak ◽  
Ricky N. S. Wong
Keyword(s):  
Cell Migration ◽  
Umbilical Vein ◽  
Cell Monolayer ◽  
Migration Assay ◽  
Migration Study ◽  
A Cell ◽  
Multiple Sample ◽  
Underlying Mechanisms ◽  
Umbilical Vein Endothelial Cells

Cell migration plays a key role in both normal physiological and pathological conditions. The study of cell migration and its underlying mechanisms is of great significance in various fields of research, including basic biology and pharmaceutical development. The cell migration or scratch wounding assay is an easy and economical in vitro method that allows researchers to assess a large number of testing compounds. Even though this simple assay has been used for decades, researchers are still trying to modify such experimental protocols and wounding devices. In this study, an 8-channel mechanical “wounder” was designed for performing a cell migration assay, particularly in a 96-well culture plate format. With special designs of a guiding bar and adjustable pins for use with disposable pipette tips, this wounder confined the scratch area within the center of each well to ensure a perfect contact between the pins and the well surface. As a result, this mechanical wounder produces a uniform denudation of a cell monolayer in a 96-well plate with a wound size of around 600 µm. Using this improved wounding device, the effects of epidermal growth factor and DL-α-difluoromethylornithine on the reepithelialization of rat intestinal epithelial cells (IEC-6) and serum on the wound recovery of human umbilical vein endothelial cells were demonstrated. This wounder facilitates cell migration study and can be applicable for multiple sample analysis.

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