RRC ID 6769
著者 Takata M, Maniwa Y, Doi T, Tanaka Y, Okada K, Nishio W, Ohbayashi C, Yoshimura M, Hayashi Y, Okita Y.
タイトル Double-layered collagen gel hemisphere for cell invasion assay: successful visualization and quantification of cell invasion activity.
ジャーナル Cell Commun Adhes
Abstract Although various methods for collagen gel-based cell invasion assays have been described, there continues to be a need for a simpler and more objective assay. Here, we describe an easy-to-prepare double-layered collagen gel hemisphere (DL-CGH) system that satisfies these requirements, and we demonstrate the advantages of this new system for visualizing cell movements during invasion. DL-CGH consists of a central core collagen layer surrounded by an outer cover collagen layer. A droplet of collagen I solution (containing cells to be examined) naturally forms a small hemisphere on the bottom of the culture dish. After this central core layer gels, a second droplet is placed atop the first gel, encapsulating it completely. The hemisphere is submerged in the medium and cultured. The invasive activity of cells that infiltrate from the inner to the outer layer can be evaluated optically. Using this in vitro system, we measured the inhibitory effect of E-cadherin expression on cancer cell invasion. DL-CGH also allowed visualization of interactions between invading cancer cells and the stroma. Cancer cells, which lack the proteases required for direct entrance into the three-dimensional collagen matrix, were seen to slip like amoebas through matrix gaps generated by the pericellular proteolytic activity of fibroblasts. [Supplementary materials are available for this article. Go to the publisher's online edition of Cell Communication and Adhesion for the following free supplemental resources: Movies 1-3; 4a and b].
巻・号 14(4)
ページ 157-67
公開日 2007-10-1
DOI 10.1080/15419060701557859
PII 783395031
PMID 17957531
MeSH Adenocarcinoma / metabolism Adenocarcinoma / pathology Biological Assay* Cadherins / antagonists & inhibitors Cadherins / genetics Cadherins / metabolism Cell Adhesion* Cell Movement* Collagen / chemistry* Collagen / metabolism Extracellular Matrix / chemistry* Extracellular Matrix / metabolism Gels / chemistry* Gels / metabolism Humans Hyaluronan Receptors / chemistry Hyaluronan Receptors / genetics Hyaluronan Receptors / metabolism Lung Neoplasms / metabolism Lung Neoplasms / pathology Matrix Metalloproteinase Inhibitors Matrix Metalloproteinases / metabolism Neoplasm Invasiveness / pathology* RNA, Small Interfering / pharmacology Stromal Cells / metabolism Stromal Cells / pathology Tumor Cells, Cultured
IF 2.5
引用数 9
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY CELL BIOLOGY
リソース情報
ヒト・動物細胞 WI-38(RCB0704)