Cardiovascular disease continues to be a leading cause of death worldwide motivating the need for models of cardiac function in both healthy and pathological conditions for basic science and clinically translational research. Cardiomyocytes (CMs) derived from human induced pluripotent stem cells provide a source for developing in vitro cardiac models, however current in vitro culture strategies and analysis techniques provide only a portion of the necessary means to fully characterize functionality. Specifically, there is a need for additional techniques to quantify and understand the contractile behavior of CMs as they interact collectively with each other and their surroundings. The following chapters describe the utility of an engineered culture platform that enables full field mechanical analysis resulting in new findings regarding CM behavior in healthy and disease models. Using a tailorable, biologically relevant platform, the influence of substrate properties and extracellular matrix (ECM) characteristics are explored. Digital Image Correlation (DIC) is used, and additional mechanical analysis tools were developed to extend the utility of the analysis pipeline. The platform was used to examine interactions between cardiac fibroblasts (CFs) and CMs, as well as the interactions of these cells with the ECM. iPSC-CFs remodel and produce aligned ECM as well as increasing contractile strain when in co-culture with iPSC-CMs. When seeded on decellularized ECM, the contractile strain was found to be higher for iPSC-CMs in co-culture with iPSC-CFs, but there was no significant difference between ECM conditions. iPSC-CMs maintained spatial organization of their contractions in co-culture with iPSC-CFs. The functionality of this platform as a disease model was then demonstrated, first as a model of catecholaminergic polymorphic ventricular tachycardia (CPVT) using iPSC-CMs from a CPVT patient (RyR2-H2464D mutation) and a healthy familial control. The maximum contractile strain was found to be consistently higher in iPSC-CMs derived from the patient compared to the familial control across three different substrate stiffnesses. Additionally, the patient cell line had a statistically significantly slower intrinsic contraction rate than the control. A hypertrophic cardiomyopathy (HCM) disease model was created using CRISPR/Cas9 modified iPSCs deficient in the cMyBP-C. The model recapitulated the increased contractile function of CMs with a homozygous knockout prior to hypertrophic remodeling. The utility of the platform was further demonstrated by evaluating the response to stimuli such as substrate stiffness and patterned features and analyzing contractile kinetics. These combined results highlight the utility of the platform as an in vitro cardiovascular model and will allow for further understanding of the interplay of genetics, environment, and genotype-phenotype relationships.

Tissue engineering integrates knowledge and tools from biological sciences and engineering for tissue regeneration. A challenge for tissue engineering is to identify appropriate cell sources. The recent advancement of stem cell biology provides enormous opportunities to engineer stem cells for tissue engineering. The impact of stem cell technology on tissue engineering will be revolutionary.This book covers state-of-the-art knowledge on the potential of stem cells for the regeneration of a wide range of tissues and organs, including cardiovascular, musculoskeletal, neurological and skin tissues. The technology platforms for studying and engineering stem cells, such as hydrogel and biomaterials development, microfluidics system and microscale patterning, are also illustrated. Regulatory challenges and quality control for clinical translation are also detailed. This book provides an comprehensive update on the advancement in the field of stem cells and regenerative medicine, and serves as a valuable resource for both researchers and students.

The book covers multifarious aspects of stem cell-based therapy for cardiovascular diseases. In addition to stem cells from different sources for cell-based therapy, it covers stem cell organoids and stem cell-derived exosomes in regenerative medicine. The book also encompasses advances in state-of-the-art infrastructure to improve the maturation aspects of pluripotent stem cells-derived cardiomyocytes using a novel scaffold-based cell culture system for cell delivery in experimental animal models and clinical settings. Besides the use of mesenchymal stem cells, the book includes chapters on the use of cardiac progenitor cells (CPCs), microtissue implantation, use of PSCs for valvulopathies, application of de-cellularized organ arrays as natural scaffolds for cardiac tissue engineering, use of epicardial stem cells, and skeletal myoblasts in cell-based therapy for myocardial regeneration. Besides the cell-based therapy approach, the book also reviews the stem cell-derived exosomes, their characteristics, and engineering strategies to enhance their therapeutic potential via targeting and drug loading and use in disease models. Additionally, the book also discusses the latest research on injectable hydrogels for cardiovascular regeneration and how hydrogel-based delivery protects the cells and their retention post-engraftment in the heart, a problem, which significantly reduces the efficacy of cell-based therapy.

Cellular and Molecular Pathobiology of Cardiovascular Disease focuses on the pathophysiology of common cardiovascular disease in the context of its underlying mechanisms and molecular biology. This book has been developed from the editors' experiences teaching an advanced cardiovascular pathology course for PhD trainees in the biomedical sciences, and trainees in cardiology, pathology, public health, and veterinary medicine. No other single text-reference combines clinical cardiology and cardiovascular pathology with enough molecular content for graduate students in both biomedical research and clinical departments. The text is complemented and supported by a rich variety of photomicrographs, diagrams of molecular relationships, and tables. It is uniquely useful to a wide audience of graduate students and post-doctoral fellows in areas from pathology to physiology, genetics, pharmacology, and more, as well as medical residents in pathology, laboratory medicine, internal medicine, cardiovascular surgery, and cardiology. - Explains how to identify cardiovascular pathologies and compare with normal physiology to aid research - Gives concise explanations of key issues and background reading suggestions - Covers molecular bases of diseases for better understanding of molecular events that precede or accompany the development of pathology

Cardiac tissue engineering enables the generation of functional human cardiac tissue using cells in combination with biocompatible materials. Human pluripotent stem cell (hPSC)-derived cardiomyocytes provide a cell source for cardiac tissue engineering; however, their immaturity limits their potential applications. Here we sought to study the effect of mechanical conditioning and electrical pacing on the maturation of hPSC-derived cardiac tissues. In the first part of the study, cardiomyocytes derived from human induced pluripotent stem cells (hIPSCs) were used to generate collagen-based bioengineered human cardiac tissue. Engineered tissue constructs were subject to different stress and electrical pacing conditions. This engineered human myocardium exhibits Frank-Starling curve-type force-length relationships. After 2 weeks of static stress conditioning, the engineered myocardium demonstrated at least 10-fold increase in contractility and tensile stiffness, greater cell alignment, and a 1.5-fold increase in cell size and cell volume fraction within the constructs. Stress conditioning also increased sarco-endoplasmic reticulum calcium transport ATPase 2 (SERCA2) expression. When electrical pacing was combined with static stress conditioning, the tissues showed an additional 2-fold increase in force production, tensile stiffness, and contractility, with no change in cell alignment or cell size, suggesting maturation of excitation-contraction coupling. Supporting this notion, we found expression of RYR2 and SERCA2 further increased by combined static stress and electrical stimulation. These studies demonstrate that electrical pacing and mechanical stimulation promote both the structural and functional maturation of hiPSC-derived cardiac tissues. In the second part of the study, cardiovascular progenitor (CVP) cells derived from hPSC were used as the input cell population to generate engineered tissues. The effects of a 3-D microenvironment and mechanical stress on differentiation and maturation of human cardiovascular progenitors into myocardial tissue were evaluated. Compared to 2-D culture, the unstressed 3-D environment increased cardiomyocyte numbers and decreased smooth muscle numbers. Additionally, 3-D culture suppressed smooth muscle cell maturation. Mechanical stress conditioning further improved cardiomyocyte maturation. Cyclic stress-conditioning increased expression of several cardiac markers, like beta-myosin and cTnT, and the tissue showed enhanced force production. This 3-D system has facilitated understanding of the effect of mechanical stress on the differentiation and morphogenesis of distinct cardiovascular cell populations into organized, functional human cardiovascular tissues. In conclusion, we were able to create a complex engineered human cardiac tissue with both stem cell-derived cardiomyocytes and CVP cells. We showed that how environmental stimulations like mechanical stress, electrical pacing, and 3-D culturing can affect the maturation and specification of cells within the engineered cardiac tissues. The study paves our way to further apply these engineered cardiac tissues to other in vitro and in vivo usages like drug testing, clinical translation, and disease modeling.

During the past decade, a wide range of scientific disciplines have adopted the use of adipose-derived stem/stromal cells (ASCs) as an important tool for research and discovery. In Adipose-Derived Stem Cells: Methods and Protocols, experts from the field, including members of the esteemed International Federation of Adipose Therapeutics and Science (IFATS), provide defined and established protocols in order to further codify the utilization of these powerful and accessible cells. With chapters organized around approaches spanning the discovery, pre-clinical, and clinical processes, much of the emphasis is placed on human ASC, while additional techniques involving small and large animal species are included. As a volume in the highly successful Methods in Molecular BiologyTM series, the detailed contributions include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Adipose-Derived Stem Cells: Methods and Protocols serves as a vital reference text for experienced researchers as well as new students on the path to further exploring the incredible potential of ASCs.

In stem cell research there are several key methods that, once mastered, can be extremely powerful. These methods enable you to rigorously test hypotheses, compare results to "gold standards," and may even spur improvements to existing protocols. This book describes numerous methods to derive, manipulate, target, and prepare stem cells for clinical use. The methods described here help you derive and test human embryonic stem cells, analyze bone marrow stem cell function in vitro and in vivo, image a stem cell transplant, cryopreserve stem cells and differentiate stem cells using microscale tec.