Engineering Strategies for Regenerative Medicine considers how engineering strategies can be applied to accelerate advances in regenerative medicine. The book provides relevant and up-to-date content on key topics, including the interdisciplinary integration of different aspects of stem cell biology and technology, diverse technologies, and their applications. By providing massive amounts of data on each individual, recent scientific advances are rapidly accelerating medicine. Cellular, molecular and genetic parameters from biological samples combined with clinical information can now provide valuable data to scientists, clinicians and ultimately patients, leading to the development of precision medicine. Equally noteworthy are the contributions of stem cell biology, bioengineering and tissue engineering that unravel the mechanisms of disease, regeneration and development. - Considers how engineering strategies can accelerate novel advances in regenerative medicine - Takes an interdisciplinary approach, integrating different aspects of research, technology and application - Provides up-to-date coverage on this rapidly developing area of medicine - Presents insights from an experienced and cross-disciplinary group of researchers and practitioners with close links to industry

​This book reviews the latest biotechnological advances with pluripotent stem cells, exploring their application in tissue engineering and medicinal chemistry. Chapters from expert contributors cover topics such as the production of transgene-free induced pluripotent stem cells (iPSCs), expansion, controlled differentiation and programming of pluripotent stem cells, and their genetic instability. Particular attention is given to the application of the pluripotent stem cells for vascularision of engineered tissue and for drug screening. This book will appeal to researchers working in regenerative medicine and drug discovery, and to bioengineers and professionals interested in stem cell research.

Human embryonic and induced pluripotent stem cells (collectively known as pluripotent stem cells or hPSCs) can be a renewable source of cardiomyocytes for treating heart diseases which are leading causes of morbidity and mortality. Recent advances in the generation of patient-specific induced PSCs greatly improve the chances of clinical success. Yet, robust methods are still under development for producing specific cardiac muscle cell-types across different hPSC lines having varying lineage-specific differentiation propensities. This requires a detailed understanding of signaling pathways and differentiation mechanisms in context of several ambient conditions such as cell epigenetic profile, endogenous signaling, and cell differentiation state. Therapeutic realization also hinges on the transplantation of sufficient numbers of cells (approximately 1. 5x109 per damaged myocardium [1]) pointing to the need for pertinent large-scale bioprocesses. In this study a novel, hormone-free, xeno-free medium is developed that is devoid of signaling activity. Since contemporary xeno-free formulations contain agents with signaling activity (such as growth hormone, retinoic acid, corticosterone, LiCl, ascorbic acid) that are reported to interfere with cardiac differentiation, proper assessment of signaling mechanisms is not feasible. Therefore a factorial design consisting of 73 conditions is employed a develop a medium comprising of specific combination of basal medium, iron-carrier (holo-transferrin), reducing agent (selenium), and a novel cocktail to increase cell growth and viability (termed XFM cocktail) containing optimized levels of lipids, aminoacids and vitamins. The medium developed here is utilized to investigate signaling mechanisms governing cardiomyocyte differentiation of hPSCs. The simple formulation of the medium offers significant economic advantages over other xeno-free media for a large scale differentiation process. Finally, since the medium is free from xenogeneic and allogeneic agents (such as albumin), clinical translation is feasible preventing the risk of transferring animal-derived pathogens and non-native proteins that can trigger adverse foreign-body immune reactions and possible implant rejection. This is the first report of accurate assessment of majority of physiologically-relevant signaling pathways elucidating their individual roles in cardiac differentiation as well as formation of closely-related lineages. By employing a design-of-experiment approach, an efficient path of hPSC-to-cardiomyocyte differentiation is discovered comprising of two steps. A synergistic combination of BMP and Wnt3A treatment with endogenous FGF and Nodal signaling is determined for obtaining an intermediate progenitor-like stage termed mesoderm-oriented primitive streak (MePS) that significantly increases differentiation efficiency. Adjustment of treatment concentration and duration during the first step and appropriate time of starting the second step allows for efficient conversion of MePS cells to cardiomyoctes in a total of 6 days of differentiation. This optimization renders the method highly reproducible that translates across the most diverse set of hPSC lines with varied lineage-specific differentiation propensities. The efficiency of hPSC-to-cardiomyocyte conversion is >90% and the cell yield is the highest reported yet in the literature providing 23 cardiomyocytes per stem cell seeded or 0. 9 million cardiomyocytes/cm2. First beating is seen as early as day 6 of differentiation and by day 8 there is interconnected and synchronized beating activity with majority of cells beating in each dish. By day 16 of differentiation, formation of bi-nucleated cells is observed indicating maturation toward adult-like cardiomyocytes. Distribution of cardiac cell-types is determined by electrophysiological measurements to be ~60% ventricular and ~40% atrial and sinoatrial node. Investigation of molecular mechanisms in the first stage reveals a crosstalk between BMP and Wnt pathways providing novel scientific information regarding mechanisms governing efficient cardiomyocyte differentiation. In the next stage, pathways are elucidated that selectively pattern primitive streak cells to different mesoderm derivatives such as smooth muscle, hematopoietic, blood and cardiac. Initial results demonstrate increased cardiac gene expression and modulation of atrial and ventricular markers providing avenues for fine-tuning mesoderm into different cardiac cell-types and possibly for increasing yield of differentiation. The outcome of this thesis is expected to facilitate the development of stem cell-based therapies for the heart.

In this book, experts in the field express their well-reasoned opinions on a range of complex, clinically relevant issues across the full spectrum of cell and gene therapies with the aim of providing trainee and practicing hematologists, including hematopoietic transplant physicians, with information that is relevant to clinical practice and ongoing research. Each chapter focuses on a particular topic, and the concise text is supported by numerous working tables, algorithms, and figures. Whenever appropriate, guidance is provided regarding the availability of potentially high-impact clinical trials. The rapid evolution of cell and gene therapies is giving rise to numerous controversies that need to be carefully addressed. In meeting this challenge, this book will appeal to all residents, fellows, and faculty members responsible for the care of hematopoietic cell transplant patients. It will also offer a robust, engaging tool to aid vital activities in the daily work of every hematology and oncology trainee.

To achieve cardiac regeneration using pluripotent stem (iPS) cells, researchers must understand iPS cell generation methods, cardiomyocyte differentiation protocols, cardiomyocyte characterization methods, and tissue engineering. This book presents the current status and future possibilities in cardiac regeneration using iPS cells. Written by top r

Cardiac Tissue Engineering: Methods and Protocols presents a collection of protocols on cardiac tissue engineering from pioneering and leading researchers around the globe. These include methods and protocols for cell preparation, biomaterial preparation, cell seeding, and cultivation in various systems. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Cardiac Tissue Engineering: Methods and Protocols highlights the major techniques, both experimental and computational, for the study of cardiovascular tissue engineering.