Embryonic Stem Cells Differentiation and Pluripotent Alternatives Edited by Michael S. Kallos EMBRYONIC STEM CELLS – DIFFERENTIATION AND PLURIPOTENT ALTERNATIVES Edited by Michael S. Kallos INTECHOPEN.COM Embryonic Stem Cells - Differentiation and Pluripotent Alternatives http://dx.doi.org/10.5772/1789 Edited by Michael S. Kallos Contributors Erhard Bieberich, Guanghu Wang, Balaji M. M. Rao, Prasenjit Sarkar, Rubens Camargo Siqueira, Mariusz Z. Ratajczak, Dong-Myung Shin, Janina Ratajczak, Magda Kucia, Keiji Miyazawa, Kyoko Kawasaki, Ronald A. Li, Ellen Poon, Chi-wing Kong, Shin Kawamata, Noemi Fusaki, Naoki Nishishita, Suk-Ying Tsang, Iek-Chi Lo, Chun-Kit Wong, Maria Elisabetta Ruaro, Jelena Ban, Vincent Torre, David Warburton, Gianni Carraro, Laura Perin, Orquidea H. Garcia, Roger De Filippo, Kazutoshi Takahashi, Koji Tanabe, Nicole I. I. Zur Nieden, Kevin C. Keller, Diego Franco, Amelia Aranega, Estefania Lozano-Velasco, Sohail Ahmed, Hui Theng Gan, Chen Sok Lam, Srinivas Ramasamy, Oz Pomp, Peter Oettgen, Katriina Aalto-Setälä, Ville J. Kujala, Mari Pekkanen-Mattila, Irina Kerkis, Mirian A.F. Hayashi, Nelson Foresto Lizier, Antonio Carlos Cassola, Alexandre Kerkis, Lygia V. Pereira, Candace Kerr, Siddharth Gupta, Angelo All, Francois Berthiaume, Timothy J. Maguire, Martin L. Yarmush, Nir I. Nativ, Mehdi A. Ghodbane, Tammy Laberge, Herman S. Cheung, Laszlo Nagy, Zoltan Simandi, Toshio Miki, Hiroshi Imai, Sandeep Goel, Mariz Vainzof, Camila De Freitas F. Almeida, Danielle Ayub-Guerrieri, Raymond C.B. Ching-Bong Wong, Ellen L. Smith, Peter J. Donovan © The Editor(s) and the Author(s) 2011 The moral rights of the and the author(s) have been asserted. All rights to the book as a whole are reserved by INTECH. 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Notice Statements and opinions expressed in the chapters are these of the individual contributors and not necessarily those of the editors or publisher. No responsibility is accepted for the accuracy of information contained in the published chapters. The publisher assumes no responsibility for any damage or injury to persons or property arising out of the use of any materials, instructions, methods or ideas contained in the book. First published in Croatia, 2011 by INTECH d.o.o. eBook (PDF) Published by IN TECH d.o.o. Place and year of publication of eBook (PDF): Rijeka, 2019. IntechOpen is the global imprint of IN TECH d.o.o. Printed in Croatia Legal deposit, Croatia: National and University Library in Zagreb Additional hard and PDF copies can be obtained from orders@intechopen.com Embryonic Stem Cells - Differentiation and Pluripotent Alternatives Edited by Michael S. Kallos p. cm. ISBN 978-953-307-632-4 eBook (PDF) ISBN 978-953-51-6521-7 Selection of our books indexed in the Book Citation Index in Web of Science™ Core Collection (BKCI) Interested in publishing with us? Contact book.department@intechopen.com Numbers displayed above are based on latest data collected. For more information visit www.intechopen.com 4,100+ Open access books available 151 Countries delivered to 12.2% Contributors from top 500 universities Our authors are among the Top 1% most cited scientists 116,000+ International authors and editors 120M+ Downloads We are IntechOpen, the world’s leading publisher of Open Access books Built by scientists, for scientists Meet the editor Dr. Michael S. Kallos, PhD, PEng is an Associate Professor of Chemical Engineering in the Schulich School of En- gineering, and Associate Director of the Pharmaceutical Production Research Facility (PPRF), both at the Univer- sity of Calgary. He is also the Director of the Biomedical Engineering Graduate Program and the Associate Direc- tor of the Center for Bioengineering Research and Educa- tion (CBRE), both at the University of Calgary. His research interests include stem cell bioprocess design, bioreactors, tissue engineering and mass trans- fer. He was awarded the Alberta Science and Technology (ASTech) “Leader of Tomorrow” Award in 2002, a Faculty of Engineering Service Award in 2005, and was named one of Calgary’s Top 40 Under 40 in 2009. He has also won numerous Teaching Awards from his peers and students, including “Professor of the Year” and “Outstanding Excellence in Teaching”. Contents Preface X III Part 1 General Differentiation 1 Chapter 1 Role of Signaling Pathways and Epigenetic Factors in Lineage Determination During Human Embryonic Stem Cell Differentiation 3 Prasenjit Sarkar and Balaji M. Rao Chapter 2 Bioactive Lipids in Stem Cell Differentiation 33 Erhard Bieberich and Guanghu Wang Chapter 3 Retinoid Signaling is a Context-Dependent Regulator of Embryonic Stem Cells 55 Zoltan Simandi and Laszlo Nagy Part 2 Neural and Retinal Differentiation 79 Chapter 4 Pluripotent Stem Cells as an In Vitro Model of Neuronal Differentiation 81 Irina Kerkis, Mirian A. F. Hayashi, Nelson F. Lizier, Antonio C. Cassola, Lygia V. Pereira and Alexandre Kerkis Chapter 5 Characterization of Embryonic Stem (ES) Neuronal Differentiation Combining Atomic Force, Confocal and DIC Microscopy Imaging 99 Maria Elisabetta Ruaro, Jelena Ban and Vincent Torre Chapter 6 Oligodendrocyte Fate Determination in Human Embryonic Stem Cells 119 Siddharth Gupta, Angelo All and Candace Kerr Chapter 7 Stem-Cell Therapy for Retinal Diseases 135 Rubens Camargo Siqueira X Contents Part 3 Cardiac and Other Myogenic Differentiation 149 Chapter 8 Transcriptional Networks of Embryonic Stem Cell-Derived Cardiomyogenesis 151 Diego Franco, Estefania Lozano-Velasco and Amelia Aránega Chapter 9 Human Pluripotent Stem Cells in Cardiovascular Research and Regenerative Medicine 169 Ellen Poon, Chi-wing Kong and Ronald A. Li Chapter 10 Human Pluripotent Stem Cell-Derived Cardiomyocytes: Maturity and Electrophysiology 185 Ville Kujala, Mari Pekkanen-Mattila and Katriina Aalto-Setälä Chapter 11 Maintenance Of Calcium Homeostasis in Embryonic Stem Cell-Derived Cardiomyocytes 205 Iek Chi Lo, Chun Kit Wong and Suk Ying Tsang Chapter 12 Myogenic Differentiation of ES Cells for Therapies in Neuromuscular Diseases: Progress to Date 227 Camila F. Almeida, Danielle Ayub-Guerrieri and Mariz Vainzof Part 4 Endothelial Differentiation 243 Chapter 13 Dissecting the Signal Transduction Pathway that Directs Endothelial Differentiation Using Embryonic Stem Cell Derived Vascular Progenitor Cells 245 Kyoko Kawasaki and Keiji Miyazawa Chapter 14 Endothelial Differentiation of Embryonic Stem Cells 267 Peter Oettgen Part 5 Hepatic Differentiation 277 Chapter 15 Stem Cells for HUMAN Hepatic Tissue Engineering 279 N.I. Nativ, M.A. Ghodbane, T.J. Maguire, F. Berthiaume and M.L. Yarmush Chapter 16 Hepatic Differentiation of Human Embryonic and Induced Pluripotent Stem Cells for Regenerative Medicine 303 Toshio Miki Part 6 Osteogenic Differentiation 321 Chapter 17 Osteogenesis from Pluripotent Stem Cells: Neural Crest or Mesodermal Origin? 323 Kevin C. Keller and Nicole I. zur Nieden Contents X I Part 7 Pluripotent Alternatives – Induced Pluripotent Stem Cells (iPSCs) 349 Chapter 18 The Past, Present and Future of Induced Pluripotent Stem Cells 351 Koji Tanabe and Kazutoshi Takahashi Chapter 19 New Techniques in the Generation of Induced Pluripotent Stem Cells 373 Raymond C.B. Wong, Ellen L. Smith and Peter J. Donovan Chapter 20 Generation of ICM-Type Human iPS Cells from CD34 + Cord Blood Cells 399 Naoki Nishishita, Noemi Fusaki and Shin Kawamata Chapter 21 Modelling of Neurological Diseases Using Induced Pluripotent Stem Cells 413 Oz Pomp, Chen Sok Lam, Hui Theng Gan, Srinivas Ramasamy and Sohail Ahmed Part 8 Pluripotent Alternatives - Other Cell Sources 431 Chapter 22 Very Small Embryonic/Epiblast-Like Stem Cells (VSELs) Residing in Adult Tissues and Their Role in Tissue Rejuvenation and Regeneration 433 Dong-Myung Shin, Janina Ratajczak, Magda Kucia and Mariusz Z. Ratajczak Chapter 23 Multipotent Dental Stem Cells: An Alternative Adult Derived Stem Cell Source for Regenerative Medicine 451 Tammy Laberge and Herman S. Cheung Chapter 24 Pluripotent Stem Cells from Testis 473 Sandeep Goel and Hiroshi Imai Chapter 25 Amniotic Fluid Stem Cells 493 Gianni Carraro, Orquidea H. Garcia, Laura Perin, Roger De Filippo and David Warburton Preface Embryonic stem cells have immense therapeutic potential, but for cell therapy these pluripotent cells will have to be differentiated towards cells of interest before transplantation. Controlled, robust differentiation processes will require knowledge of the signalling pathways and mechanisms which will obviously be unique for each differentiated cell type. There are a tremendous variety of approaches and techniques, and this book, Embryonic Stem Cells - Differentiation and Pluripotent Alternatives and its companion, Embryonic Stem Cells - Basic Biology to Bioengineering , serve as a snapshot of many of the activities currently underway on a number of different fronts. This book is divided into eight parts and provides examples of the many tissue types that embryonic stem cells are being pushed towards, as well as alternative sources for pluripotent stem cells. Part 1: General Differentiation Chapters 1-3 present a number of different aspects of embryonic stem cell differentiation including signalling pathways, epigenetic factors, bioactive lipids and retinoid signalling. Part 2: Neural and Retinal Differentiation Chapters 4-7 examine how neural cell types are generated from embryonic stem cells, including looking at some interesting imaging techniques and a review of stem cell therapy for retinal diseases. Part 3: Cardiac and Other Myogenic Differentiation Chapters 8-12 present a number of aspects of cardiomyocytes differentiation including detailed characterization of the differentiated cells. In addition the use of ESC-derived myogenic cells for neuromuscular diseases is discussed. Part 4: Endothelial Differentiation Chapters 13-14 describe endothelial differentiation with a focus on the signalling transduction pathways involved. Part 5: Hepatic Differentiation Chapters 15-16 examine another interesting application of embryonic stem cells for hepatic tissue engineering. X Preface Part 6: Osteogenic Differentiation Chapter 17 looks at osteogenesis and the developmental path from pluripotent cells to osteoblasts. Part 7: Pluripotent Alternatives Induced Pluripotent Stem Cells (iPSCs) Chapters 18-21 discuss induced pluripotent stem cells (iPSCs) starting with a great review of the iPSC story to date. The next chapters present new techniques to generate iPSCs including from cord blood cells. An interesting application of iPSCs in the modelling of neurological diseases is described as an example of one of the many uses of these cells. Part 8: Pluripotent Alternatives Other Cell Sources Chapters 22-25 describe alternative sources of pluripotent stem cells. These include very small embryonic/epiblast like stem cells, multipotent dental stem cells, pluripotent stem cells from testis and amniotic fluid stem cells. In the book Embryonic Stem Cells - Differentiation and Pluripotent Alternatives , the story begins with a foundation upon which future therapies and uses of embryonic stem cells can be built. I would like to thank all of the authors for their valuable contributions. I would also like to thank Megan Hunt who provided me with much needed assistance and acted as a sounding board for early chapter selection, and the staff at InTech, particularly Romina Krebel who answered all of my questions and kept me on track during the entire process. Calgary, Alberta, Canada, September 2011 Michael S. Kallos Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, Alberta, Canada Department of Chemical and Petroleum Engineering, Schulich School of Engineering, University of Calgary, Alberta, Canada Part 1 General Differentiation 1 Role of Signaling Pathways and Epigenetic Factors in Lineage Determination During Human Embryonic Stem Cell Differentiation Prasenjit Sarkar and Balaji M. Rao Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA 1. Introduction Human embryonic stem cells (hESCs) are culture-adapted cells that were originally derived from the inner cell mass (ICM) of the blastocyst-stage embryo [1]. HESCs are pluripotent cells that can be propagated indefinitely in culture, while retaining the in vivo properties of ICM cells; they can give rise to all tissues of the three germ layers (ectoderm, mesoderm and endoderm). Due to their pluripotency, hESCs have been the subject of intense research since they were initially isolated in 1998. HESCs can serve as model systems to study early human development, in addition to providing a potentially unlimited source of functional tissues for use in drug evaluation and regenerative medicine. Nevertheless, despite major advances, the exact molecular mechanisms that govern the self-renewal and differentiation of hESCs remain unclear. Indeed, a mechanistic understanding of the molecular processes regulating hESC fate can elucidate early events in human development and enable the development of protocols for efficient generation of functional tissues. Here we review the molecular mechanisms that regulate hESC fate; specifically, we focus on the role of signaling pathways and factors regulating epigenetic changes, in hESC self-renewal and lineage-specific differentiation. In hESCs, as in embryos, differentiation is triggered by developmental cues such as morphogens or cytokines that are present in the extracellular space. These morphogens or cytokines bind to their cognate plasma membrane-bound receptors and activate specific signaling pathways inside the cell. Activation of signaling pathways involves a sequence of phosphorylation events that eventually result in the regulation of specific transcription factors. These transcription factors, in turn, can recruit other co-factors and directly cause transcription of downstream genes. Furthermore, transcription factors can recruit histone modifying and chromatin remodeling enzymes to reshuffle the epigenetic structure, such that pluripotency genes become inaccessible for transcription and are repressed, whereas lineage-specific genes become accessible and are activated. This sequence of events finally leads to expression of lineage-specific proteins such as transcription factors and structural proteins, causing a morphological change in the cell. Also, pluripotency associated transcription factors and other pluripotency-associated genes are permanently repressed, thereby completing the process of differentiation. Thus, the process of differentiation is a Embryonic Stem Cells – Differentiation and Pluripotent Alternatives 4 rather complex cascade of events, controlled by signaling pathways, transcription factors, epigenetic factors and lineage-specific proteins. While significant understanding of each of these functional groups (i.e., signaling pathways, transcription factors, epigenetic factors and lineage-specific proteins) has been gathered in isolation, very little is known about the interactions amongst these groups, particularly in the context of hESC differentiation. In part, interactions amongst these groups confer lineage specificity to the process of differentiation and mediate the development of specific tissues upon exposure of hESCs to certain morphogens. In this review, we focus on the role of signaling pathways, transcription factors and epigenetic factors in the context of lineage-specific differentiation of hESCs and summarize the various links between these groups. Our goal is to present a mechanistic overview of the sequence of molecular events that regulate the differentiation of hESCs along various lineages. 2. The signaling pathways As briefly described earlier, the self-renewal and differentiation of hESCs is governed by several developmental cues. The most well known among these are cytokines that trigger specific signaling pathways. These extracellular ligands initiate signaling through interactions with ligand-specific cell surface receptors. Receptor-binding typically results in association of multiple receptor subunits and activation of the kinase domains of receptors or other receptor-bound effector proteins. This triggers a sequence of phosphorylation events involving various other proteins, finally resulting in the activation or inhibition of transcription factors. These transcription factors in turn are directly responsible for activating or repressing their target genes. Thus, a group of signaling pathways is usually responsible for modulating gene expression in hESCs, leading to control of the transcriptome, the proteome and ultimately cellular physiology. In this section, we summarize key signaling pathways that have been implicated in the maintenance of undifferentiated hESCs and their lineage-specific differentiation. 2.1.1 The transforming growth factor- β pathway The transforming growth factor (TGF- ) pathway is well known for its involvement in embryonic development and patterning, as well as in epithelial-to-mesenchymal transformations and carcinogenesis [2-3]. This pathway (extensively reviewed elsewhere [2, 4-5]) is divided into two branches: the Activin/Nodal branch and the Bone Morphogenetic Protein (BMP) branch. The Activin/Nodal pathway is activated by ligands such as Activin, Nodal and TGF 1. These ligands bind to their Type II receptor, which then recruits the Type I receptor. The Type II receptor phosphorylates the intracellular domain of the Type I receptor, creating a binding site for SMAD2 and SMAD3 transcription factors. Upon binding, SMAD2/3 is phosphorylated by the Type I receptor, leading to subsequent dissociation of SMAD2/3. Phosphorylated SMAD2/3 then associates with SMAD4 and can enter the nucleus to modulate gene expression. The BMP branch is activated by the BMP ligands; binding of BMP ligands results in phosphorylation of the type I receptor by the type II receptor, and subsequent intracellular binding and phosphorylation of SMAD1/5/8. Phosphorylated SMAD1/5/8 forms a complex with SMAD4 and subsequently enters the nucleus. Unlike ligands in the Activin/Nodal branch, the BMPs have high affinities for the type I receptor and bind weakly to the type II receptor; Activin/Nodal bind with high