EMBRYONICSTEMCELLS– BASICBIOLOGYTO BIOENGINEERING EditedbyMichaelS.Kallos Embryonic Stem Cells – Basic Biology to Bioengineering Edited by Michael S. Kallos Published by InTech Janeza Trdine 9, 51000 Rijeka, Croatia Copyright © 2011 InTech All chapters are Open Access articles distributed under the Creative Commons Non Commercial Share Alike Attribution 3.0 license, which permits to copy, distribute, transmit, and adapt the work in any medium, so long as the original work is properly cited. After this work has been published by InTech, authors have the right to republish it, in whole or part, in any publication of which they are the author, and to make other personal use of the work. Any republication, referencing or personal use of the work must explicitly identify the original source. 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 articles. 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. Publishing Process Manager Romina Krebel Technical Editor Teodora Smiljanic Cover Designer Jan Hyrat Image Copyright Knorre, 2011. Used under license from Shutterstock.com First published August, 2011 Printed in Croatia A free online edition of this book is available at www.intechopen.com Additional hard copies can be obtained from orders@intechweb.org Embryonic Stem Cells – Basic Biology to Bioengineering, Edited by Michael S. Kallos p. cm. ISBN 978-953-307-278-4 free online editions of InTech Books and Journals can be found at www.intechopen.com Contents Preface IX Part 1 Challenges and Possibilities – From New Cell Lines to Alternative Uses of Cryopreserved Embryos 1 Chapter 1 Embryonic Stem Cells for Therapies – Challenges and Possibilities 3 Ronne Wee Yeh Yeo and Sai Kiang Lim Chapter 2 Derivation and Characterization of New hESC Lines from Supernumerary Embryos, Experience from Turkey 19 Zafer Nihat Candan and Semra Kahraman Chapter 3 Cryopreserved Embryos: A Catholic Alternative to Embryonic Stem Cell Research and Adoption 33 Peter A. Clark Part 2 Methods, Tools and Technologies for Embryonic Stem Cell Culture, Manipulation and Clinical Application 47 Chapter 4 Bioprocess Development for the Expansion of Embryonic Stem Cells 49 Megan M. Hunt, Roz Alfred, Derrick E. Rancourt, Ian D. Gates and Michael S. Kallos Chapter 5 Small-Scale Bioreactors for the Culture of Embryonic Stem Cells 73 Allison Van Winkle, Ian D. Gates and Michael S. Kallos Chapter 6 Synthetic Surfaces for Human Embryonic Stem Cell Culture 89 Andrei G. Fadeev and Zara Melkoumian Chapter 7 Efficient Integration of Transgenes and Their Reliable Expression in Human Embryonic Stem Cells 105 Kenji Sakurai, Miho Shimoji, Kazuhiro Aiba and Norio Nakatsuji VI Contents Chapter 8 Embryonic Stem Cells: Introducing Exogenous Regulators into Embryonic Stem Cells 123 Yong-Pil Cheon Chapter 9 Functional Control of Target Single Cells in ES Cell Clusters and Their Differentiated Cells by Femtoinjection 149 Hideaki Matsuoka, Mikako Saito and Hisakage Funabashi Chapter 10 From Pluripotency to Early Differentiation of Human Embryonic Stem Cell Cultures Evaluated by Electron Microscopy and Immunohistochemistry 171 Janus Valentin Jacobsen, Claus Yding Andersen, Poul Hyttel and Kjeld Møllgård Part 3 Applications of Embryonic Stem Cells in Research and Development 191 Chapter 11 Methods to Generate Chimeric Mice from Embryonic Stem Cells 193 Kun-Hsiung Lee Chapter 12 Embryonic Stem Cells in Toxicological Studies 213 Carmen Estevan, Andrea C. Romero, David Pamies, Eugenio Vilanova and Miguel A. Sogorb Chapter 13 Teratomas Derived from Embryonic Stem Cells as Models for Embryonic Development, Disease, and Tumorigenesis 231 John A. Ozolek and Carlos A. Castro Part 4 Pluripotency and Molecular Biology of Embryonic Stem Cells 263 Chapter 14 Illuminating Hidden Features of Stem Cells 265 Gideon Grafi, Rivka Ofir, Vered Chalifa-Caspi and Inbar Plaschkes Chapter 15 Signaling Pathways in Mouse Embryo Stem Cell Self-Renewal 283 Leo Quinlan Chapter 16 Building a Pluripotency Protein Interaction Network for Embryonic Stem Cells 305 Patricia Ng and Thomas Lufkin Chapter 17 Profile of Galanin in Embryonic Stem Cells and Tissues 321 Maria-Elena Lautatzis and Maria Vrontakis Chapter 18 Rho-GTPases in Embryonic Stem Cells 333 Michael S. Samuel and Michael F. Olson Contents VII Chapter 19 Cripto-1: At the Crossroads of Embryonic Stem Cells and Cancer 347 Nadia Pereira Castro, Maria Cristina Rangel, Tadahiro Nagaoka, Hideaki Karasawa, David S. Salomon and Caterina Bianco Chapter 20 Molecular Mechanisms Underlying Pluripotency and Lineage Commitment – The Role of GSK-3 369 Bradley W. Doble, Kevin F. Kelly and James R. Woodgett Part 5 Lessons from Development 389 Chapter 21 Embryonic Stem Cells and the Germ Cell Lineage 391 Cyril Ramathal, Renee Reijo Pera and Paul Turek Chapter 22 Techniques and Conditions for Embryonic Germ Cell Derivation and Culture 425 Maria P De Miguel, Candace L Kerr, Pilar López-Iglesias and Yago Alcaina Chapter 23 Pluripotent Gametogenic Stem Cells of Asexually Reproducing Invertebrates 449 Valeria V. Isaeva Preface TheisolationandcultureofhumanembryonicstemcellsbyThomsoninthelate1990s hasacceleratedaparadigmshiftinmedicinethatwasstartedmuchearlierbyTilland McCulloch in the early 1960s with the discovery of the first stem cells in mice. The burgeoning field of regenerative medicine will ultimately transform mo dern human healthcarefromamolecule‐basedfocus,whichservestoalleviatesymptoms,toacell andtissuebasedfocuswhichhasthepromiseofactuallyrestoringfunction.Although the potential is enormous, the road is long and there are certainly many milestones alongtheway. Thisbook, Embryonic StemCells‐ Basic Biologyto Bioengineeringand its companion,EmbryonicStemCells‐DifferentiationandPluripotentAlternatives,serveasa snapshotofmanyoftheactivitiescurrentlyunderwayonanumberofdifferentfronts. This book is divided into five parts and provides a foundation upon which future therapiesandus esofembryonicstemcellscanbebuilt. Part1:ChallengesandPossibilities‐FromNewCellLinestoAlternativeUsesofCryopreserved Embryos Chapters1‐3 offerabroad overviewof some ofthechallenges in bringingembryonic stemcellbasedmedicinetotheclinic,aswellasacasestu dyofthederivationofnew embryonicstemcelllines,andanalternativetotheuseofcryopreservedembryos. Part2:Methods,ToolsandTechnologiesforEmbryonicStemCellCulture,Manipulationand ClinicalApplication Chapters 4‐10 present a wide variety of tools and technologies ranging from large‐ scale bi oreactors to scaled‐down bioreactor arrays and synthetic surfaces that can be used for embryonic stem cell culture. In addition, methods for introducing foreign genesintoembryonicstemcellsandcontrollinggeneexpressionaredescribed.Lastly, the use of imaging is presented as a tool to measure pluripotency and early differentiation. Part3:Applic ationsofEmbryonicStemCellsinResearchandDevelopment Chapters11‐13 presentmethods togenerate chimericmice foruse in research,and in addition,describetheuseofembryonicstemcellsintoxicologicalstudiesandtheuse X Preface of teratomas derived from embryonic stem cells as models for early development, disease,andtumorigenesis. Part4:PluripotencyandMolecularBiologyofEmbryonicStemCells Chapters14‐20describeourunderstandingofpluripotencyaswellassomeofthekey molecules involved in regulating not only pluripotency but cancer and early embryonictiss ues. Part5:LessonsfromDevelopment Chapters 21‐23 examine the knowledge we have gained from studying embryonic germ cells and pluripotent gametogenic stem cells of asexually reproducing invertebrates. In the book EmbryonicStem Cells‐Differentiation andPluripotent Alternatives, thestory continueswithasampleofsomeofthestu diescurrentlyunderwaytoderiveneural, cardiac,endothelial,hepaticandosteogeniclineages.Inaddition,inducedpluripotent stem cells are introduced and other unique sources of pluripotent stem cells are explored. I would like tothank allof the authorsfor their valuable contributions. Iwould also liketo thankMegan Huntwho providedme with muc h neededassistanceand 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 entireprocess. Calgary,Alberta,Canada,July2011 MichaelS.Kallos PharmaceuticalProductionR esearchFacility(PPRF), SchulichSchoolofEngineering,UniversityofCalgary,Alberta Canada DepartmentofChemicalandPetroleumEngineering, SchulichSchoolofEngineering,UniversityofCalgary,Alberta, Canada [...]... strategy, Choo et al has raised antibodies against undifferentiated hESCs (Choo et al., 2008) and identified an antibody that was cytotoxic against hESCs by oncosis This antibody was an IgM that recognizes podocalyxin-like protein-1(PODXL) hESCs that were treated with mAB 84 did not form teratoma when transplanted into SCID mice even after 18-24 weeks Therefore, there are viable technologies to remove or reduce... progenitor E-RoSH cells, ERoSHK cells also exhibit enhanced activity in biochemical pathways that are also highly characteristic of beta cells such as the pentose phosphate pathway, clathrin-mediated endocytosis and PPAR signaling (T S Chen et al., 2010) Importantly, transplantation of ERoSHK cells in hyperglycemic streptozotocintreated mice reverses the hyperglycemia and removal of the transplanted... of evolutionary selection, driven by environmental pressures rather than by conscious design Therefore, instead of mapping experimental protocols on to the known developmental pathways of pancreatic endocrine cells, they proposed that conscious design may be a less circuitous route to arrive at the same end-point However, in lieu of known developmental pathways, there is no obvious source to guide and... formation by human embryonic stem cells Nat Biotechnol, Vol 27, No 3, (Mar), pp (281-287) Burns, C.J., Persaud, S.J., & Jones, P.M (2004) Stem cell therapy for diabetes: do we need to make beta cells? J Endocrinol, Vol 183, No 3, (Dec), pp (437-443) Byrne, J.A., Pedersen, D.A., Clepper, L.L., Nelson, M., Sanger, W.G., Gokhale, S., Wolf, D.P., & Mitalipov, S.M (2007) Producing primate embryonic stem cells... Teh, B.J., Luo, R., Li, G., & Lim, S.K (2010) Delineating biological pathways unique to embryonic stem cell-derived insulin-producing cell lines from their noninsulin-producing progenitor cell lines Endocrinology, Vol 151, No 8, (Aug), pp (3600-3610) Chidgey, A.P., Layton, D., Trounson, A., & Boyd, R.L (2008) Tolerance strategies for stemcell-based therapies Nature, Vol 453, No 7193, (May 15), pp (330-337)... Ang, S.N., Fong, W.J., Chin, A., Lo, J., Zheng, L., Hentze, H., Philp, R.J., Oh, S.K., & Yap, M (2008) Selection against undifferentiated human embryonic stem cells by a cytotoxic antibody recognizing podocalyxin-like protein1 Stem Cells, Vol 26, No 6, (Jun), pp (1454-1463) Conlon, F.L., Lyons, K.M., Takaesu, N., Barth, K.S., Kispert, A., Herrmann, B., & Robertson, E.J (1994) A primary requirement for . antibody that was cytotoxic against hESCs by oncosis. This antibody was an IgM that recognizes podocalyxin-like protein-1(PODXL). hESCs that were treated with mAB 84 did not form teratoma when