Description
Humana Press Metastasis Research Protocols (Methods in Molecular Medicine, 58) 1st Editon 2010 Softbound by Susan A. Brooks, Udo Schumacher
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. 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.