In Volume I, Analysis of Cells and Tissues, we presented a range of protocols aimed at mapping and analyzing the expression of various molecules of pot- tial interest in metastasis research and for examining their production at the genetic level. In this second volume of metastasis research protocols, we move to the level of living cells and tissues and present methodologies applicable to examining metastatic behavior in vitro and in whole animal models. The methods described in the first section of this volume concentrate on the separation of cell lines with high and low metastatic potential, including the genetic modification of cell lines. The assay systems to test defined aspects of the metastatic cascade are then described in Part II and include cell migration assays, assays for matrix degrading enzymes, basement membrane degrading assays, adhesion assays, and assays of angiogenesis. The role of the specific elements of the metastatic cascade assayed in each of these systems in turn must of course be put into perspective relative to their roles in entire living organisms.

Includes supplementary material: sn.pub/extras

Klappentext

Although ninety percent of fatal cancer cases involve the spread of a primary tumor, the formation of metastases is still a poorly understood, complex process and a significant problem in the treatment of cancer patients. In Metastasis Research Protocols, leading international investigators describe in detail the key methods needed to investigate why and how metastasis occurs. Volume II of this two-volume set, Analysis of Cell Behavior In Vitro and In Vivo, presents a comprehensive collection of established and leading-edge techniques for examining metastatic behavior in vitro and in whole animal models. Methods are provided for the separation of cell lines with high and low metastatic potential, along with assay systems to test defined aspects of the metastatic cascade. These systems include cell migration assays, assays for matrix degradation enzymes, basement membrane degradation assays, adhesion assays, and assays of angiogenesis. The detailed descriptions of animal models for metastasis cover the use of immunodeficient animals, syngenic and transgenic models, and orthotopic models of metastasis. The first volume of this set, Analysis of Cells and Tissues, focuses on the analysis and mapping of molecules produced by cells and tissues, and on the molecular biology underlying their expression.
Comprehensive and authoritative, the two volumes of Metastasis Research Protocols constitute a gold-standard collection of readily reproducible methods for understanding the metastatic cascade-first at the cellular and molecular levels, then at the level of the whole organism-responsible for the spread of cancer and for developing novel strategies to combat its spread.

Inhalt
Selection/Segregation Cell Populations for in Vitro and in Vivo Assays of Metastatic Behavior.- Cell Separations by Flow Cytometry.- Immunomagnetic Cell Separation.- Genetic Modification of Cell Lines to Enhance Their Metastatic Capability.- In Vitro Assays of Metastatic Behavior.- Cell Aggregation Assays.- Cell Migration and the Boyden Chamber.- Quantification of Cell Motility.- In Vitro Invasion Assay Using Matrigel®.- Membrane Invasion Culture System.- Collagen Invasion Assay.- Chick Heart Invasion Assay.- Adhesion of Tumor Cells to Matrices and Endothelium.- Assessment of Angiogenic Factors The Chick Chorioallantoic Membrane Assay.- Methods to Evaluate the Formation and Stabilization of Blood Vessels and Their Role in Tumor Growth and Metastasis.- Galectin-3 Binding and Metastasis.- Animal Models of Metastasis.- Basic Principles for the Study of Metastasis Using Animal Models.- The Chick Embryo Metastasis Model.- GeneTransfection for Metastasis Research Using Animal Models.- Theoretical Considerations in Using Animal Models of Metastasis and Brief Methodology for In Vivo Colorectal Cancer Models in SCID and Nude Mice.- Xenograft Models in Immunodeficient Animals.- Xenograft Models in Immunodeficient Animals.- Syngeneic Murine Metastasis Models.- Transgenic Animal Models.- Clinically Accurate Orthotopic Mouse Models of Cancer.- Dissection of Tumor and Host Cells from Metastasized Organs for Testing Gene Expression Directly Ex Vivo.- Green Fluorescent Protein for Metastasis Research.