干细胞之家www.stemcell8.cn ←点击进入 Tissue Engineering Essentials for Daily Laboratory Work W. W. Minuth, R. Strehl, K. Schumacher 干细胞之家www.stemcell8.cn ←点击进入 干细胞之家www.stemcell8.cn ←点击进入 Tissue Engineering W. W. Minuth, R. Strehl, K. Schumacher 干细胞之家www.stemcell8.cn ←点击进入 Further Titles of Interest Novartis Foundation Symposium Kay C. Dee, David A. Puleo, Rena Bizios Tissue Engineering An Introduction to Tissue- of Cartilage and Bone – Biomaterial Interactions No. 249 2002 ISBN 0-471-25394-4 2003 ISBN 0-470-84481-7 Alan Doyle, J. Bryan Griffiths (Eds.) Rolf D. Schmid, Ruth Hammelehle Cell and Tissue Culture Pocket Guide to Biotechnology for Medical Research and Genetic Engineering 2000 2003 ISBN 0-471-85213-9 ISBN 3-527-30895-4 R. Ian Freshney Michael Hoppert Culture of Animal Cells: Microscopic Techniques A Manual of Basic Technique, in Biotechnology 4th Edition 2003 ISBN 3-527-30198-4 2000 ISBN 0-471-34889-9 R. Ian Freshney, Mary G. Freshney Oliver Kayser, Rainer H. Mu¨ller (Eds.) (Eds.) Pharmaceutical Biotechnology: Culture of Epithelial Cells, Drug Discovery and Clinical 2nd Edition Applications 2002 2004 ISBN 0-471-40121-8 ISBN 3-527-30554-8 干细胞之家www.stemcell8.cn ←点击进入 Tissue Engineering Essentials for Daily Laboratory Work W. W. Minuth, R. Strehl, K. Schumacher 干细胞之家www.stemcell8.cn ←点击进入 Authors This book was carefully produced. Nevertheless, editors, authors and publisher do not warrant the Dr. Will W. Minuth, PhD information contained therein to be free of errors. Raimund Strehl, PhD Readers are advised to keep in mind that state- Karl Schumacher, M.D. ments, data, illustrations, procedural details or other items may inadvertently be inaccurate. University of Regensburg Department of Molecular and Cellular Anatomy Library of Congress Card No.: Applied for. University Street 31 93053 Regensburg British Library Cataloguing-in-Publication Data: Germany A catalogue record for this book is available from the British Library. Bibliografische Information Der Deutschen Bibliothek Translated by Die Deutsche Bibliothek lists this publication in the Deutsche Nationalbibliografie; detailed Renate FitzRoy (Chapters 6, 8) bibliographic data is available in the internet at 26 Cairnhill Gardens http://dnb.ddb.de. St Andrews, Scotland KY16 8BX ª 2005 Wiley-VCH Verlag GmbH & Co. KGaA, Nicole Heath (Chapters 1, 2, 3, 4, 7, 9, 10, Glossary) Weinheim Fischergasse 6 69117 Heidelberg All rights reserved (including those of translation in other languages). No part of this book may be Matthias Herbst (Chapter 5) reproduced in any form – nor transmitted or Markgra¨flerstr. 12 translated into a machine language without written 69126 Heidelberg permission from the publishers. Registered names, trademarks, etc. used in this book, even when not specifically marked as such, are not to be con- sidered unprotected by law. Printed in the Federal Republic of Germany. Important Note: Printed on acid-free paper. As research and clinical work are constantly ex- panding our knowledge, we would like to empha- Composition Mitterweger & Partner, Plankstadt size that when this book was written, all dosage and Printing betz-druck GmbH, Darmstadt application specifications reflected the state of the Bookbinding Großbuchbinderei J. Scha¨ffer art. However, users are strongly advised to check the GmbH & Co. KG, Gru¨nstadt instructions that come with the preparations and medical products used and use their own judge- ISBN-13: 978-3-527-31186-6 ment on dosage according to specific recommen- ISBN-10: 3-527-31186-6 dations in their own countries. Finding Literature Using Search Criteria Due to the rapid growth of information, constantly transforming our knowledge in the area of cell biology and tissue engineering , it made sense to collate a number of search criteria rather than a bibliography. Put in a medical or biological database such as PubMed or Biological Abstracts, these search criteria will lead you to the most up-to-date literature on the subject. 干细胞之家www.stemcell8.cn ←点击进入 V Preface Why this book at this time? A number of things have come together. In restructuring our lab, we needed to clear out, organize and archive. A lot of interesting material from the past was lying around that, for various reasons, was not being further investigated and thus had never been published. On inspection of the data and images, we realized that we had actually learned much more from unsuccessful experiments than from the successful ones that had seamlessly fit into the experimental design. When we came upon difficulties, we did not give up. We continually asked new questions and carried out further experiments until we came to logical explanations. In addition, we have offered many courses in cell and tissue culture as well as tissue engineering over the years, for participants from both Germany and abroad. The participants often asked interesting and fundamental questions which were insuffi- ciently or completely unanswered by previous books. To solve this problem, it was necessary to do a great deal of research in the various databanks. We have sketched, structured and worked the answers to those questions into the text as funda- mental information. Although we train students daily in microscopic anatomy, it has become increas- ingly evident to us how little is known about the development of functional tis- sues. However, it is exactly this aspect that is of particular importance for the future production of tissue constructs, from adult cells or stem cells, for use in patients. Socially interactive cell networks must be produced out of individual cells and im- planted into the patient as functional tissue, and no health risks should be added in the process. This book introduces theoretically fundamental and experimental concepts, which should open the door into the field of tissue engineering. Additionally, it should give students, technicians and young scientists a look into the fascinating world of differ- entiable cells and tissues. We must make clear that we stand at the beginning of a very exciting and future-oriented scientific development. For this reason, we must adjust ourselves to learning about the development of tissues. After sufficient experimenta- tion, and in the course of this decade, tissue engineering will change from a purely empirical to an analytically reproducible science. We will get an overview of each step in tissue development and learn to simulate it experimentally. Apart from molecular biological processes, epigenetic factors and microenvironments will also play a major 干细胞之家www.stemcell8.cn ←点击进入 VI Preface roll. In addition, we must adjust to the fact that it will not be possible to generate functional tissues with cell culture methods. Will W. Minuth, R. Strehl, K. Schumacher Regensburg, February 2003 干细胞之家www.stemcell8.cn ←点击进入 VII Contents Preface V 1 Developmental processes 1 2Cells and Tissue 4 2.1 The Cell 4 2.1.1 The Cell as a Functional Unit 4 2.1.2 Plasma Membrane 5 2.1.3 Nucleus 6 2.1.4 Mitochondria 6 2.1.5 Endoplasmic Reticulum (ER) 6 2.1.6 Golgi Apparatus 7 2.1.7 Endosomes, Lysosomes and Peroxisomes 7 2.1.8 Cytoskeleton 8 2.1.9 ECM 8 2.1.10 Cell Cycle 9 2.2 Tissue Types 10 2.2.1 Epithelia 10 2.2.1.1 Building Plans of Epithelia 11 2.2.1.2 Glands 14 2.2.1.3 Epithelia in Sensory Perception 16 2.2.2 Connective Tissue 17 2.2.2.1 Variety 18 2.2.2.2 Fat Tissue as Storage 20 2.2.2.3 Bone and Cartilage as Support Tissue 21 2.2.3 Muscle Tissue 26 2.2.3.1 Cell Movement 26 2.2.3.2 Rhythmic Contraction 28 2.2.3.3 Unconscious Contraction 29 2.2.4 Nervous System Tissue 31 2.2.4.1 Information Mediation 31 2.2.4.2 Networks and Connections 33 干细胞之家www.stemcell8.cn ←点击进入 VIII Contents 2.3 Relevance of the ECM 35 2.3.1 Components of the ECM 35 2.3.1.1 Functions of the ECM 35 2.3.1.2 Synthesis of the Collagens 37 2.3.1.3 Fibronectin 38 2.3.1.4 Laminin 39 2.3.1.5 Reticular and Elastic Fibers 39 2.3.1.6 Collagens of the Basement Membrane 39 2.3.1.7 FACIT Collagens 40 2.3.1.8 Proteoglycans 40 2.3.2 Interactions between the Cell and the ECM 41 2.3.2.1 Adhesion and the ECM 41 2.3.2.2 Proliferation and the ECM 41 2.3.2.3 Differentiation and the ECM 42 2.3.2.4 Apoptosis and the ECM 43 2.3.3 Signal Transduction 43 2.3.3.1 Modulation of the Cell– Matrix Interaction 43 2.3.3.2 The ECM and Cell Binding 44 2.3.3.3 Signals to the Inner Cell 47 2.3.3.4 The ECM and Long-term Contact 48 2.3.4 Matricellular Proteins 51 2.3.4.1 Thrombospondin 52 2.3.4.2 Tenascin C 52 2.3.4.3 Osteopontin 52 2.3.4.4 SPARC 53 2.4 Emergence of Tissue 53 2.4.1 Germ Layers and Ground Tissue 53 2.4.1.1 Derivatives of the Ectoderm 55 2.4.1.2 Derivatives of the Mesoderm 56 2.4.1.3 Derivatives of the Entoderm 58 2.4.2 Individual Cells, Social Interactions and Functional Tissue Develop- ment 58 2.4.2.1 Differentiation from Individual Cells 59 2.4.2.2 Functional Exceptions 60 2.4.2.3 Individual Cells and Social Interactions 60 2.4.2.4 Formation of tissue 61 2.4.2.5 Individual Cell Cycles 66 2.4.2.6 Coordinated Growth 67 2.4.2.7 Competence 67 2.4.2.8 Morphogenic Factors 68 2.4.2.9 Apoptosis 69 2.4.2.10 Necrosis versus Apoptosis 71 2.4.2.11 Terminal Differentiation 71 2.4.2.12 Adaptation 72 2.4.2.13 Transdifferentiation 73 干细胞之家www.stemcell8.cn ←点击进入 Contents IX 2.4.2.14 Multifactorial Differentiation 73 2.5 Regeneration 74 2.5.1 Events Immediately after an Injury 74 2.5.2 Wound Closure 75 2.5.3 Programmed Cell Death (Apoptosis) 75 2.5.4 Cooperative Renewal 76 3 Classical Culture Methods 78 3.1 History 78 3.2 First Cultures 79 3.2.1 Culture Containers 80 3.2.1.1 Individual Culture Containers 80 3.2.1.2