Boyden chamber-based method for characterizing the distribution of adhesions and cytoskeletal structure in HT1080 fibrosarcoma cells

J. Tóvári, Krisztina Futosi, Alexandra Bartal, Eniko Tátrai, Alexandra Gacs, I. Kenessey, S. Paku

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A 2D model was previously presented that describes the gliding motility of human fibrosarcoma cells. The model was based on the observation that adhesions are present only on the outer rim of the leading lamella of the semicircular cell. The present model describes the organization of adhesions and the cytoskeleton of migrating HT1080 fibrosarcoma and LX2 hepatic stellate cells in three dimensions. The migration assays were performed in a modified Boyden chamber using fibronectin, Matrigel, or collagen I as chemoattractants. The distribution of the adhesions was analyzed by confocal laser scanning microscope, and following decoration with heavy meromyosin, the organization of actin filaments was analyzed by electron microscopy. Double labeling was performed to study the relationship of the actin and vimentin filament network in the moving cells. Vinculin containing adhesions were observed only at the front of the cell in the form of a ring while passing through a filter pore of the Boyden chamber. Actin filaments were present only below the plasma membrane, except the very tip of the leading lamella. Vimentin intermediate filaments were localized around the cell nucleus behind the actin filament-rich lamella. This paper describes a model of the organization of adhesions and the cytoskeleton of migrating cells in the Boyden chamber. The model is based on the observation that adhesions are present only at the leading edge of the cell. The results extend the earlier 2D model of cell locomotion into 3D.

Original languageEnglish
Pages (from-to)509-516
Number of pages8
JournalCell Adhesion and Migration
Volume8
Issue number5
DOIs
Publication statusPublished - Sep 1 2014

Fingerprint

Fibrosarcoma
Actin Cytoskeleton
Vimentin
Cytoskeleton
Vinculin
Myosin Subfragments
Hepatic Stellate Cells
Intermediate Filaments
Chemotactic Factors
Cell Nucleus
Fibronectins
Cell Movement
Electron Microscopy
Lasers
Collagen
Cell Membrane

Keywords

  • Adhesion plaque dynamics
  • Boyden chamber
  • Cell migration
  • Fibroblast-type motility
  • Migration assay

ASJC Scopus subject areas

  • Cell Biology
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Boyden chamber-based method for characterizing the distribution of adhesions and cytoskeletal structure in HT1080 fibrosarcoma cells. / Tóvári, J.; Futosi, Krisztina; Bartal, Alexandra; Tátrai, Eniko; Gacs, Alexandra; Kenessey, I.; Paku, S.

In: Cell Adhesion and Migration, Vol. 8, No. 5, 01.09.2014, p. 509-516.

Research output: Contribution to journalArticle

@article{8b9054f825ab40799cf09a5dd84f2593,
title = "Boyden chamber-based method for characterizing the distribution of adhesions and cytoskeletal structure in HT1080 fibrosarcoma cells",
abstract = "A 2D model was previously presented that describes the gliding motility of human fibrosarcoma cells. The model was based on the observation that adhesions are present only on the outer rim of the leading lamella of the semicircular cell. The present model describes the organization of adhesions and the cytoskeleton of migrating HT1080 fibrosarcoma and LX2 hepatic stellate cells in three dimensions. The migration assays were performed in a modified Boyden chamber using fibronectin, Matrigel, or collagen I as chemoattractants. The distribution of the adhesions was analyzed by confocal laser scanning microscope, and following decoration with heavy meromyosin, the organization of actin filaments was analyzed by electron microscopy. Double labeling was performed to study the relationship of the actin and vimentin filament network in the moving cells. Vinculin containing adhesions were observed only at the front of the cell in the form of a ring while passing through a filter pore of the Boyden chamber. Actin filaments were present only below the plasma membrane, except the very tip of the leading lamella. Vimentin intermediate filaments were localized around the cell nucleus behind the actin filament-rich lamella. This paper describes a model of the organization of adhesions and the cytoskeleton of migrating cells in the Boyden chamber. The model is based on the observation that adhesions are present only at the leading edge of the cell. The results extend the earlier 2D model of cell locomotion into 3D.",
keywords = "Adhesion plaque dynamics, Boyden chamber, Cell migration, Fibroblast-type motility, Migration assay",
author = "J. T{\'o}v{\'a}ri and Krisztina Futosi and Alexandra Bartal and Eniko T{\'a}trai and Alexandra Gacs and I. Kenessey and S. Paku",
year = "2014",
month = "9",
day = "1",
doi = "10.4161/cam.28734",
language = "English",
volume = "8",
pages = "509--516",
journal = "Cell Adhesion and Migration",
issn = "1933-6918",
publisher = "Landes Bioscience",
number = "5",

}

TY - JOUR

T1 - Boyden chamber-based method for characterizing the distribution of adhesions and cytoskeletal structure in HT1080 fibrosarcoma cells

AU - Tóvári, J.

AU - Futosi, Krisztina

AU - Bartal, Alexandra

AU - Tátrai, Eniko

AU - Gacs, Alexandra

AU - Kenessey, I.

AU - Paku, S.

PY - 2014/9/1

Y1 - 2014/9/1

N2 - A 2D model was previously presented that describes the gliding motility of human fibrosarcoma cells. The model was based on the observation that adhesions are present only on the outer rim of the leading lamella of the semicircular cell. The present model describes the organization of adhesions and the cytoskeleton of migrating HT1080 fibrosarcoma and LX2 hepatic stellate cells in three dimensions. The migration assays were performed in a modified Boyden chamber using fibronectin, Matrigel, or collagen I as chemoattractants. The distribution of the adhesions was analyzed by confocal laser scanning microscope, and following decoration with heavy meromyosin, the organization of actin filaments was analyzed by electron microscopy. Double labeling was performed to study the relationship of the actin and vimentin filament network in the moving cells. Vinculin containing adhesions were observed only at the front of the cell in the form of a ring while passing through a filter pore of the Boyden chamber. Actin filaments were present only below the plasma membrane, except the very tip of the leading lamella. Vimentin intermediate filaments were localized around the cell nucleus behind the actin filament-rich lamella. This paper describes a model of the organization of adhesions and the cytoskeleton of migrating cells in the Boyden chamber. The model is based on the observation that adhesions are present only at the leading edge of the cell. The results extend the earlier 2D model of cell locomotion into 3D.

AB - A 2D model was previously presented that describes the gliding motility of human fibrosarcoma cells. The model was based on the observation that adhesions are present only on the outer rim of the leading lamella of the semicircular cell. The present model describes the organization of adhesions and the cytoskeleton of migrating HT1080 fibrosarcoma and LX2 hepatic stellate cells in three dimensions. The migration assays were performed in a modified Boyden chamber using fibronectin, Matrigel, or collagen I as chemoattractants. The distribution of the adhesions was analyzed by confocal laser scanning microscope, and following decoration with heavy meromyosin, the organization of actin filaments was analyzed by electron microscopy. Double labeling was performed to study the relationship of the actin and vimentin filament network in the moving cells. Vinculin containing adhesions were observed only at the front of the cell in the form of a ring while passing through a filter pore of the Boyden chamber. Actin filaments were present only below the plasma membrane, except the very tip of the leading lamella. Vimentin intermediate filaments were localized around the cell nucleus behind the actin filament-rich lamella. This paper describes a model of the organization of adhesions and the cytoskeleton of migrating cells in the Boyden chamber. The model is based on the observation that adhesions are present only at the leading edge of the cell. The results extend the earlier 2D model of cell locomotion into 3D.

KW - Adhesion plaque dynamics

KW - Boyden chamber

KW - Cell migration

KW - Fibroblast-type motility

KW - Migration assay

UR - http://www.scopus.com/inward/record.url?scp=84921766480&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84921766480&partnerID=8YFLogxK

U2 - 10.4161/cam.28734

DO - 10.4161/cam.28734

M3 - Article

C2 - 25482525

AN - SCOPUS:84921766480

VL - 8

SP - 509

EP - 516

JO - Cell Adhesion and Migration

JF - Cell Adhesion and Migration

SN - 1933-6918

IS - 5

ER -