AVB Training Program: Cell and Molecular Biology
Trainees have the opportunity to receive extensive training in cell and molecular biology techniques which can be applied to the study of problems in the area of atherosclerosis and vascular biology. Both predoctoral and postdoctoral participants can learn isolation and primary cell culture of all the major cell types of the vessel wall. These include human monocytes, human arterial and umbilical vein endothelial cells, rabbit aortic endothelial cells, human and rabbit aortic and venous smooth muscle cells. In addition, proficiency can be obtained in the isolation and culture of primary human, rabbit, and rat hepatocytes. Techniques for production and screening of antibody secreting hybridoma cells can be learned as well as methods for large scale production of the antibodies in vivo, as ascites fluid, and in vitro in a bioreactor. Isolated cells can be utilized to address problems relating to lipoprotein receptors, in vitro metabolism of lipoprotein components, phenotypic changes accompanying lipid loading, in vitro activity of lipolytic enzymes, expression of cell adhesion molecules, expression of and phenotypic changes induced by growth factors and immune cytokines, in vitro modeling of angiogenesis, and in vitro studies of factors regulating hypertension, and intracellular signaling.
Training can be provided for numerous routine and innovative techniques of molecular biology. Included are gene cloning and sequencing, PCR, in situ hybridization, production of stable and transient in vitro transfectants, methods of vector construction for gene delivery in vivo, production of large quantities of cloned proteins using E. coli, yeast, baculovirus, and CHO or COS expression systems, design and use of antisense DNA and RNA reagents, analysis of gene fine structure including promoter regions, and transgenic overexpression and knockout constructs. Trainees apply these techniques to the study of gene regulation in vitro and in vivo of targets important in atherosclerosis and vascular biology. Applications include examination of basal gene expression as well as regulation of gene expression by pharmacologic agents, dietary manipulation, dynamic flow, and nucleotide inhibitors. The application of these techniques to gene therapy in cardiovascular disease can also be explored. Instruction in genomic techniques such as fabrication and use of gene microarrays and in situ hybridization is available; proteomic methods such as fabrication of tissue arrays, multi-ligand immunochemical quantitation, and laser capture microscopy are taught.
Participants also can be trained to utilize a variety of animal models for their studies of atherosclerosis and vascular biology. These include the cholesterol fed rabbit, the WHHL rabbit, the spontaneously hypertensive rat, the rat and rabbit restenosis model, and transgenic mice and rabbits. Program faculty participating in providing instruction in these areas are listed below.