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Cell migration assay

A cell migration, cell technology, applied in the field of preparation and use of these compositions, compositions for transendothelial migration assays, can solve problems such as being unsuitable for high-throughput screening, complex preparation, etc.

Inactive Publication Date: 2008-09-10
GE HEALTHCARE BIO SCI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this system is complex to prepare and not suitable for high-throughput screening

Method used

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  • Cell migration assay
  • Cell migration assay
  • Cell migration assay

Examples

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[0034] Preparation of the composition

[0035] We provide detailed materials and methods used to prepare the assay system in the Examples section. Briefly, collagen is deposited in the container and solidifies to form a solid layer. Or alternatively, synthetic matrix gels that support 3D endothelial growth and cell migration can be used. Optionally, a gelatin solution is added to the collagen gel before the collagen layer solidifies. The first cell type (endothelial cells) is then placed on the solid layer and cultured to form a confluent layer of cells in contact with the solid layer. Migrated cells are then prepared and seeded on top of a confluent layer of the first cell type. Typically, the first cell type is endothelial cells, such as HUVECs. Other primary endothelial cells such as HCAEC (coronary artery endothelial cells), HMVEC (pulmonary microvascular endothelial cells) or endothelial cell lines such as SK-HEP-1 (ATCC HTB-52) can also be used. Although any migra...

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Abstract

The present invention provides compositions and methods for preparation of a three-dimensional transendothelial cell migration (TEM) assay. These compositions and methods are uniquely suited for the high throughput TEM assay, and for the analysis and identification of TEM mediators which inhibit or stimulate this process. The composition for detecting migration of cells comprises a solid layer comprising collagen gel; a first cellular layer in contact with the solid layer and comprising a first cell type; and a second cell type seeded on top of the first cellular layer. Optionally, gelatin is included in the collagen gel. A 96 well plate format is disclosed, the combination with a high throughput cellular scanner enables high throughput TEM assay.

Description

[0001] Cross References to Related Applications [0002] This application claims priority to U.S. Provisional Patent Application No. 60 / 718,057, filed September 16, 2005, and U.S. Provisional Patent Application No. 60 / 747,430, filed May 17, 2006, which disclose The contents of all are incorporated herein by reference. field of invention [0003] The present invention generally relates to methods for assaying hematodialysis. More particularly, the present invention relates to compositions for use in transendothelial migration assays, and methods of making and using these compositions. Background of the invention [0004] Migration of cells across the vascular endothelium is an important event in the pathophysiology of conditions like inflammation, arteriosclerosis and tumor metastasis. Methods for measuring cell migration in vitro have been developed over the years. The most common method uses an artificial barrier (membrane) and usually requires manual counting of migrat...

Claims

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Application Information

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IPC IPC(8): C12N5/06C12Q1/02
Inventor 於利敏赵立红A・贝莱茨基P・钱纳瓦哈拉
Owner GE HEALTHCARE BIO SCI CORP
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