Though the heart is one of the first organs to develop during embryogenesis and the physical aspects of development are well documented, little is known of the molecular mechanisms that control heart development. BMP signaling has been implicated in cardiac development both in vivo and in vitro, the initial research focused on altering this pathway. BMP signaling belongs to the signaling superfamily of transforming growth factor-b (Tgf-(beta)). Further evidence from mouse knockout studies, reveals a critical role of signaling through the Tgf-(beta) receptors in which Tgf-(beta) 3-/- mice demonstrate congenital heart defects. Tgf-(beta) signaling is typically relayed through a tetramer complex composed of two Tgf-(beta) type II and two type I (ALK5) receptors. The signaling of this tetramer has recently been identified in the differentiation of epicardial and endocardial to mesenchyme. Proceeding experiments have demonstrated that knocking ALK5 out selectively in endocardium, myocardium, or epicardium does not interfere with normal cardiac muscle development in vivo. Sridurongrit suggest that ALK5 signaling is required for smooth muscle development and vascularization of the myocardium but not cardiomyocoyte development. Therefore the role of ALK5 signaling during cardiac development is studied int two pluripotent models, mouse embryonic stem cells and human induced pluripotent stem cells (hiPS) in this research to understand the role of this pathway in cardiogenesis. Further the ultimate goals of this research is to screen small molecules and develop protocols that direct diffentiation of pluripotent stem cells to mesoderm and ultimately a cardiomyocyte fate. There are two major differentiation events that occur as a pluripotent stem cell differentiates to a terminal state. The cell begins as a pluripotent cell that can give rise to all somatic cell types as this cell differentiates it enters multipotent stage. Multipotent cells become partially programmed and can give rise to only certain somatic fates. These multipotent progenitors will ultimately give rise to structured tissue composed of specific somatic cell types. However, the molecular pathways that control differentiation to specific somatic fates remain poorly understood. The focus of this research is to explore these pathways using small molecule inhibitors to better understand the internal cell signaling that controls cardiogenesis. The research presented in this paper occurs in two major stages. First the experiments focus on developing protocols that can induce pluripotent stem cells to give rise to mesoderm, the germ layer from which cardiomyocytes are derived. Secondly, small molecules are screened to understand their ability to drive this mesoderm to a cardiomyocyte fate. Exploring these pathways, that control cardiogenesis, is essential if stem cells are to provide a supply of primary cardiomycotes to better understand human cardiac physiology and the affect potential drugs will have on their function. Heart disease remains the number one cause of death in the developed world. Therefore there is not only a need to develop novel molecules that can assuage cardiac disease but there is also a need to understand how these diseases develop. hiPS have the potential to fulfill both these needs. These cells can be derived directly from patients with specific cardiac afflictions. By controlling the differentiation of these disease derived pluripotent cells, researchers will be able to track physical and chemical changes in cardiomyocyte development that ultimately lead to a diseased phenotype. This creates a powerful tool to study new molecules and cardiac disease. Screening of small molecules that alter the diseased phenotype of these patients will further understanding of chemical modulation of cardiomyocytes and the ability of potential drugs to mitigate disease. This research has the potential to ultimately lead to patient specific therapeutics in the treatment of heart disease.

This volume covers all aspects of embryonic stem cell differentiation, including mouse embryonic stem cells, mouse embryonic germ cells, monkey and human embryonic stem cells, and gene discovery.* Early commitment steps and generation of chimeric mice* Differentiation to mesoderm derivatives* Gene discovery by manipulation of mouse embryonic stem cells

Human pluripotent stem cells such as human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC) with their unique developmental plasticity hold immense potential as cellular models for drug discovery and in regenerative medicine as a source for cell replacement. While hESC are derived from a developing embryo, iPSC are generated with forced expression of key transcription factors to convert adult somatic cells to ESC-like cells, a process termed reprogramming. Using iPSC overcomes ethical issues concerning the use of developing embryos and it can be generated from patient-specific or disease-specific cells for downstream applications. Pluripotent Stem Cells: Methods and Protocols highlights the best methods and systems for the entire work flow. Divided into four convenient sections, topics include a focus on producing iPSC from diverse somatic sources, media systems for expanding ESC and iPSC with detailed protocols for directed differentiation into specific lineages, commonly used cellular and molecular characterization methods , and the potential application of labeled stem cells with specific methods for cloning, gene delivery and cell engineering. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Pluripotent Stem Cells: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essential cellular feature.

Stem cell science has the potential to impact human reproductive medicine significantly - cutting edge technologies allow the production and regeneration of viable gametes from human stem cells offering potential to preciously infertile patients. Written by leading experts in the field Stem Cells in Reproductive Medicine brings together chapters on the genetics and epigenetics of both the male and female gametes as well as advice on the production and regeneration of gene cells in men and women, trophoblasts and endometrium from human embryonic and adult stem cells. Although focussing mainly on the practical elements of the use of stem cells in reproductive medicine, the book also contains a section on new developments in stem cell research. The book is essential reading for reproductive medicine clinicians, gynecologists and embryologists who want to keep abreast of practical developments in this rapidly developing field.

Since the first successful isolation and cultivation of human embryonic stem cells at the University of Wisconsin, Madison in 1998, there has been high levels of both interest and controversy in this area of research. This book provides a concise overview of an exciting field, covering the characteristics of both human embryonic stem cells and pluripotent stem cells from other human cell lineages. The following chapters describe state-of-the-art differentiation and characterization of specific ectoderm, mesoderm and endoderm-derived lineages from human embryonic stem cells, emphasizing how these can be used to study human developmental mechanisms. A further chapter discusses genetic manipulation of human ES cells. The concluding section covers therapeutic applications of human ES cells, as well as addressing the ethical and legal issues that this research have raised.