Chemical and mechanical cues are known to guide the differentiation of pluripotent stem cells (PSCs) towards specific cell-types; a process known as directed differentiation. Typically, directed differentiation protocols to guide PSCs to specific cell types have focused on chemical cues. For example, lung epithelial directed differentiation protocols mimic the chemical cues observed in lung organogenesis. The products of current directed differentiation protocols are typically unorganized cellular aggregates that are often limited by their inability to generate specific cell types reliably. This suggests further improvements to these protocols are necessary for reliable cell manufacturing. Introducing developmentally relevant mechanical signals offers a potentially novel strategy to improve the predictability and reduce the heterogeneity of current directed differentiation protocols. To assess the feasibility of this concept we use polymeric substrates to guide the self-assembly of human PSC-derived lung progenitors into developmentally relevant sized tubes and assess the impact of defined architecture on differentiation. Specifically, day 17 ventralized lung progenitors, dual positive for proximal and distal lung markers SOX2 and SOX9, respectively, were cultured in polymeric tubes with diameters of 100μm and allowed to self-assemble into cellular tubes. Cells cultured in tubes were observed to become primarily SOX9+ as compared to cells in flat cultures that remained dual positive. Cells from tube cultures were retrieved and exposed to proximal or distal airway inducing conditions. Our data show that cells from flat culture are capable of expressing proximal and distal markers under proximal and distal conditions, respectively. However, cells retrieved from tubes were able to express distal markers under distal conditions but not proximal markers under proximal conditions. Our results suggest that modulation of tension in the presence of canonical WNT signaling by culturing the dual positive cells in the 100μm tubes resulted in loss of SOX2 expression. Overall, this thesis suggests that the use of a bioinspired mechanical microenvironment to organize and control the differentiation of developing lung epithelial populations during directed differentiation is a tractable strategy to control cellular differentiation in vitro.

Pluripotent stem cells have distinct characteristics: self-renewal and the potential to differentiate into various somatic cells. In recent years, substantial advances have been made from basic science to clinical applications. The vast amount knowledge available makes obtaining concise yet sufficient information difficult, hence the purpose of this book. In this book, embryonic stem cells, induced pluripotent stem cells, and mesenchymal stem cells are discussed. The book is divided into five sections: pluripotency, culture methods, toxicology, disease models, and regenerative medicine. The topics covered range from new concepts to current technologies. Readers are expected to gain useful information from expert contributors.

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

​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.

With this valuable practical guide, three members of the Harvard Stem Cell Institute have compiled and edited the definite handbook for the exciting new field of human embryonic stem cell research. The editors have gathered protocols from scientists with extensive reputation and expertise, describing and comparing currently used techniques for the culture of human stem cells and discussing the strengths and weaknesses of the different approaches. Human Embryonic Stem Cells: The Practical Handbook contains the first centralised collection of methods used in human embryonic stem cell biology. The book covers the derivation of human stem cell lines, the obtaining of cells from human stem cell banks, the culturing and characterisation of the cells, and the differentiation of the cells in vitro and in vivo. Lastly, almost all of these protocols can also be used for analyzing and manipulating induced pluripotency iPS stem cells. This allows an even greater number of opportunities for those interested in purusing work in pluripotent stem cells, disease modelling, and other aspects of basic regenerative medicine research. The novel and useful focus of this book sets it apart from other available books: Compares and evaluates the protocols used in leading laboratories working on human embryonic stem cells Centred solely on practical protocols for human (not mouse) embryonic stem cell research Includes extensive troubleshooting sections Addresses the different proclivities and behaviours of individual human embryonic cell lines Contains techniques currently known only to a small number of specialised laboratories worldwide This handbook represents an essential source of up-to-date practical information for all cell and developmental biologists working with human embryonic stem cells or wishing to enter the field. It is also essential reading for clinical researchers in areas such as diabetes, cardiovascular disease, and neurological diseases. Praise from the reviews: "...a highly readable and useful book... A notable feature of the book is its air of openness and honesty... This book... will help many to navigate the uncharted waters of human embryonic stem cell biology." BRITISH SOCIETY FOR CELL BIOLOGY "... the imaginative solutions in this book can inspire us to get past our most frustrating limitations." CELL STEM CELL "... the richness in the details of each protocol presented will certainly encourage more scientists to begin studies of Human pluripotent stem cells..." REGENERATIVE MEDICINE "In this fast-moving field, this [handbook] will help drive advances of more and more researchers." DIFFERENTIATION "...a valuable resource for seasoned and novice researchers... an excellent addition to the reference collection of any medical library or research laboratory." THE AMERICAN MEDICAL ASSOCIATION

This book covers the identification and role of endogenous lung stem cells in health and disease, particularly the most recent advances. In addition, it discusses the rapidly growing field of stem cells and cell therapy as it relates to lung biology and disease as well as ex vivo lung bioengineering. Such approaches may provide novel therapeutic approaches for lung diseases. Human pluripotent stem cell differentiation to model the pulmonary epithelium and vasculature is also discussed. World-recognized scientists who specialize in studying both the lung epithelium and pulmonary vasculature contribute the chapters. Topics covered include: stem cell niches in the lung, the role of progenitor cells in fibrosis and asthma, iPSC in modeling lung disease, vascular repair by endothelial progenitor cells and circulating fibrocytes in pulmonary vascular remodeling. This volume of the Stem Cell Biology and Regenerative Medicine series is essential reading for researchers and clinicians interested in stem cells, lung biology and regenerative medicine. It is also an invaluable resource for advanced students studying cell biology, regenerative medicine and lung physiology.

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.