A LABORATORY GUIDE TO THE MAMMALIAN EMBRYO

A LABORATORY GUIDE TO THE MAMMALIAN EMBRYO

Edited by

David K. Gardner Michelle Lane Andrew J. Watson

1
2004

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Oxford New York Auckland Bangkok Buenos Aires Cape Town Chennai Dar es Salaam Delhi Hong Kong Istanbul Karachi Kolkata Kuala Lumpur Madrid Melbourne Mexico City Mumbai Nairobi So Paulo Shanghai Taipei Tokyo Toronto

Copyright 2004 by Oxford University Press

Published by Oxford University Press, Inc., 198 Madison Avenue, New York, New York 10016 www.oup.com Oxford is a registered trademark of Oxford University Press All rights reserved. No part of this publication may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without the prior permission of Oxford University Press. Library of Congress Cataloging-in-Publication Data A laboratory guide to the Mammalian Embryo / edited by David K. Gardner, Michelle Lane, Andrew J. Watson. p. ; cm. Includes bibliographical references and index. ISBN 0-19-514226-8 1. Embryology, ExperimentalLaboratory manuals. 2. Embryology, HumanLaboratory manuals. [DNLM: 1. EmbryoLaboratory Manuals. 2. Cells, CulturedLaboratory Manuals. 3. Reproductive Techniques, AssistedLaboratory Manuals. QS 625 L123 2003] I. Gardner, David K. II. Lane, Michelle. III. Watson, Andrew J. (Andrew John), 1959 QL961 .L33 2003 571.8'619dc2l 2003000329

9 8 7 6 5 4 3 2 1 Printed in the United States of America on acid-free paper

Preface

The 1990s saw an exponential increase in research on the preimplantation mammalian embryo. Basic research on laboratory animals has been paralleled with increases in the use of assisted breeding techniques in both domestic animals and humans. Concomitantly, there have been improvements in the technology used to study the early embryo. New biochemical, microscopic, and genetic techniques have been developed and applied at the single-cell level. It is therefore the scope of this book to describe in detail such new developments and to aid the researcher in applying such technology in the laboratory by providing detailed protocols that augment the theoretical aspects of each chapter. Rather than focusing on a single species, the chapters cover embryos from rodents, domestic animals, and primates, including humans. The techniques and procedures described include updates of established procedures such as in vitro maturation, in vitro fertilization, and embryo culture, together with newer research areas such as the establishment of human embryonic stem cells, gamete transplantation, cloning, and genetic analysis. Although no single text can cover the entire field of mammalian embryology, we have endeavored to provide todays laboratory workers with a practical guide to many of the techniques used in modern embryology. This book should therefore serve as a useful tool for both students and experienced researchers alike.

ACKNOWLEDGMENTS
The editors thank all of the authors for their valuable contributions and Angela Hunsinger for her assistance with the preparation of this text. This book is dedicated to Jackie, Glen, and Patricia for their tireless support and understanding.

Contents

Contributors ix Abbreviations xiii 1. Follicle Development in Vitro 3 Norah Spears, Evelyn E. Telfer, and Alison A. Murray 2. Preparation of Gametes, in Vitro Maturation, in Vitro Fertilization, and Embryo Recovery and Transfer 24 Michelle Lane and David K. Gardner 3. Culture of the Mammalian Preimplantation Embryo 41 David K. Gardner and Michelle Lane 4. Assessment of Preimplantation Embryo Development and Viability 62 Denny Sakkas, Michelle Lane, and David K. Gardner 5. Micromanipulation of Gametes 76 Gianpiero D. Palermo, Queenie V. Neri, Takumi Takeuchi, Lucinda L. Veeck, and Zev Rosenwaks 6. Micromanipulation: Biopsy 99 Joyce C. Harper and Alpesh Doshi 7. Analysis of Intracellular Ions in Embryos: pH and Calcium 125 Michelle Lane, Jay M. Baltz, and David K. Gardner 8. Assessment of Nutrient Uptake, Metabolite Production, and Enzyme Activity 139 David K. Gardner and Michelle Lane 9. Metabolic Pathway Activity 154 Don Rieger 10. Confocal Imaging of Structural Molecules in Mammalian Gametes 165 Calvin Simerly, Ricardo Moreno, Joo Ramalho-Santos, Laura Hewitson, and Gerald Schatten

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CONTENTS

11. Fluorescent in situ Hybridisation for Detection of Aneuploidy in Single Human Blastomeres for Preimplantation Genetic Diagnosis 184 Leeanda Wilton 12. Assessment of Ploidy, Telomere Length, and Telomerase Activity in Oocytes and Embryos 193 W. Allan King, Dean H. Betts, Wafa Bureau, and Parvathi K. Basrur 13. Microscale RNA Isolation from Mammalian Embryos 217 Christine Wrenzycki, Andrew J. Watson, and Michele Calder 14. Relative mRNA Transcript Abundance in Early Embryos by Reverse Transcription-Polymerase Chain Reaction 225 Paul A. De Sousa, and Andrew J. Watson 15. Characterization of Novel Genes during Early Development by Application of Differential Display RTPCR 237 David R. Natale and Andrew J. Watson 16. Two-Dimensional Protein Gel Database Analysis of Embryos, Oocytes, and Oocyte-Associated Granulosa Cells 247 Keith E. Latham, Gula Nourjanova, Karen Wigglesworth, and John J. Eppig 17. Antisense Disruption of Gene Expression in the Preimplantation Embryo 258 Lisa C. Barcroft, D. Holstead Jones-Taggart, and Andrew J. Watson 18. Gene Subtraction and Analysis 270 Marc-Andr Sirard and Claude Robert 19. Analysis of Apoptosis in the Preimplantation Embryo 279 Daniel R. Brison, Anthony D. Metcalfe, Debra J. Bloor, Helen R. Hunter, Gerard Brady, and Susan J. Kimber 20. Cytoplasmic Signaling and Cell Cycle Control in the Mouse Egg and Embryo 298 D. Page Baluch, Christine M. Pauken, and David G. Capco 21. Human Embryonic Stem Cells 313 Susan E. Lazendorf, Catherine A. Boyd, and Diane L. Wright 22. Spermatogonial Transplantation 334 Makoto C. Nagano 23. Somatic Cell Nuclear Transfer 352 Paul De Sousa, William Ritchie, Andras Dinnyes, Tim King, Lesley Paterson, and Ian Wilmut 24. Cryobiology: Slow Freezing Vitrification of Embryos 375 Tetsunori Mukaida and Magosaburo Kasai Index 391

Contributors

Jay M. Baltz Departments of Obstetrics and Gynecology (Division of Reproductive Medicine), and Cellular and Molecular Medicine University of Ottawa, Canada D. Page Baluch Molecular and Cellular Biology Program School of Life Sciences Arizona State University Tempe, Arizona, USA Lisa C. Barcroft Department of OB/GYN and Physiology The University of Western Ontario London, Ontario, Canada Parvathi K. Basrur Department of Biomedical Sciences Ontario Veterinary College University of Guelph Guelph, Ontario, Canada Dean H. Betts Department of Biomedical Sciences Ontario Veterinary College University of Guelph Guelph, Ontario, Canada Debra J. Bloor School of Biological Sciences University of Manchester Manchester, UK

Catherine A. Boyd The Jones Institute Eastern Virginia Medical School Norfolk, Virginia USA Gerard Brady School of Biological Sciences University of Manchester Manchester, UK Daniel R. Brison Department of Reproductive Medicine St Marys Hospital, Manchester and School of Biological Sciences University of Manchester Manchester, UK Wafa Bureau Department of Biomedical Sciences Ontario Veterinary College University of Guelph Guelph, Ontario, Canada Michele Calder Departments of Obstetrics and Gynaecology and Physiology The University of Western Ontario London, Ontario, Canada David G. Capco Molecular and Cellular Biology Program School of Life Sciences Arizona State University Tempe, Arizona, USA ix

CONTRIBUTORS
Paul A. De Sousa Roslin Institute Roslin, Midlothian, UK Andras Dinnyes Roslin Institute Roslin, Midlothian, UK Alpesh Doshi Department of Obstetrics and Gynaecology University College London and Assisted Conception Unit University College Hospital London, UK John J. Eppig The Jackson Laboratory Bar Harbor, Maine, USA David K. Gardner Colorado Center for Reproductive Medicine Englewood, Colorado, USA Joyce C. Harper Department of Obstetrics and Gynaecology University College London and Assisted Conception Unit University College Hospital London, UK Laura Hewitson Department of Obstetrics, Gynecology and Reproductive Sciences University of Pittsburgh School of Medicine, and The Pittsburgh Development Center Pittsburgh, Pennsylvania, USA Helen R. Hunter Department of Reproductive Medicine St Marys Hospital Manchester, UK D. Holstead Jones-Taggart Ontario Cancer Institute Princess Margaret Hospital University of Toronto Toronto, Ontario, Canada Magosaburo Kasai Laboratory of Animal Science College of Agriculture Kochi University Nankoku, Kochi, Japan Susan J. Kimber School of Biological Sciences University of Manchester Manchester, UK Tim King Roslin Institute Roslin, Midlothian, UK W. Allan King Department of Biomedical Sciences Ontario Veterinary College University of Guelph Guelph, Ontario, Canada Michelle Lane Colorado Center for Reproductive Medicine Englewood, Colorado, USA Susan E. Lanzendorf Department of Obstetrics and Gynecology Washington University in St. Louis Missouri, USA Keith E. Latham The Fels Institute for Cancer Research and Molecular Biology and the Department of Biochemistry Temple University School of Medicine Philadelphia, Pennsylvania, USA Anthony D. Metcalfe School of Biological Sciences University of Manchester Manchester, UK Ricardo Moreno Unit of Reproduction and Development Physiology Department Pontifical Catholic University of Chile Santiago, Chile Tetsunori Mukaida Hiroshima HART Clinic Ohtemachi, Nakaku, Hiroshima, Japan Alison A. Murray Department of Biomedical Sciences University of Edinburgh Edinburgh, UK Makoto C. Nagano Department of Obstetrics and Gynecology Royal Victoria Hospital McGill University Montreal, Quebec, Canada

CONTRIBUTORS xi
David R. Natale Department of Biochemistry and Molecular Biology University of Calgary Calgary, Alberta, Canada Queenie V. Neri The Center for Reproductive Medicine and Infertility Weill Medical College of Cornell University New York, New York, USA Gula Nourjanova Cold Spring Harbor Gel Laboratory Cold Spring Harbor, New York, USA Gianpiero D. Palermo The Center for Reproductive Medicine and Infertility Weill Medical College of Cornell University New York, New York, USA Lesley Paterson Roslin Institute Roslin, Midlothian, UK Christine M. Pauken Molecular and Cellular Biology Program Department of Biology Arizona State University Tempe, Arizona, USA Joo Ramalho-Santos Department of Zoology University of Coimbra Coimbra, Portugal Don Rieger Animal Biotechnology Embryo Laboratory Department of Biomedical Sciences University of Guelph Guelph, Ontario, Canada William Ritchie Roslin Institute Roslin, Midlothian, UK Claude Robert Centre de Recherche en Biologic de la Reproduction Dpartement des Sciences Animales Universit Laval Qubec, Canada Zev Rosenwaks The Center for Reproductive Medicine and Infertility Weill Medical College of Cornell University New York, New York, USA Denny Sakkas Department of Obstetrics and Gynecology Yale University School of Medicine New Haven, Connecticut, USA Gerald Schatten Development of Obstetrics, Gynecology and Reproductive Sciences University of Pittsburgh School of Medicine, and The Pittsburgh Development Center, and The Magee-Womens Research Institute Pittsburgh, Pennsylvania, USA Calvin Simerly Department of Obstetrics, Gynecology and Reproductive Sciences University of Pittsburgh School of Medicine, and The Pittsburgh Development Center Pittsburgh, Pennsylvania, USA Marc-Andr Sirard Centre de Recherche en Biologic de la Reproduction Dpartement des Sciences Animales Universit Laval Qubec, Canada Norah Spears Department of Biomedical Sciences University of Edinburgh Edinburgh, UK Takumi Takeuchi The Center for Reproductive Medicine and Infertility Weill Medical College of Cornell University New York, New York, USA Evelyn E. Teller Institute of Cell and Molecular Biology University of Edinburgh Edinburgh, UK Lucinda L. Veeck The Center for Reproductive Medicine and Infertility Weill Medical College of Cornell University New York, New York, USA

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CONTRIBUTORS
Andrew J. Watson Department of Obstetrics and Gynecology and Physiology University of Western Ontario London, Ontario, Canada Leeanda Wilton Melbourne IVF Victoria, Australia Christine Wrenzycki Department of Biotechnology Institut fur Tierzucht und Tierverhalten (FAL) Mariensee, Neustadt, Germany Diane L. Wright Vincent Reproductive Medicine and IVF Massachusetts General Hospital Boston, Massachusetts, USA

Karen Wigglesworth The Jackson Laboratory Bar Harbor, Maine, USA

Ian Wilmut Roslin Institue Roslin, Midlothian, UK

Abbreviations

acetyl-CoA ART bp BSA cGMP CHAPS DAPI DMEM DMSO DNP dpm DTT EDTA EtOH FBS FISH FITC FSH GnRH hCG HEPES hMG HSA HTF IBMX ICM ICSI i.d. IVF IVM kb KSOM

acetyl coenzyme A assisted reproductive techniques basepair bovine serum albumin cyclic guanidine 5'-monophosphate [(cholamidopropyl)dimethylammonio]-l-propanesulfonate 4',6-diamidino-2-phenylindole dihydrochloride Dulbeccos modified Eagles medium dimethylsulfoxide dinitrophenol disintegrations per minute dithiothreitol ethylenediaminetetraacetic acid ethanol fetal bovine serum flurorescence in situ hybridization fluorescein isothiocyanate follicle-stimulating hormone gonadotropin-releasing hormone human chorionic gonadotropin N-(2-hydroxyethyl)piperazine-N'-ethanesulfonic acid human menopausal gonadotropin human serum albumin human tubal fluid medium 3-Isobutyl-l-methylxanthine inner cell mass intracytoplasmic sperm injection inner diameter in vitro fertilization in vitro maturation kilobase modified simplex optimized medium with elevated potassium xiii

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ABBREVIATIONS
LH MEM MOPS mw o.d. PAGE PBS PCR PMSF PMSG PVA PVP SDS SSC SSCP RT-PCR TAE TBE TBS TC TE TEMED TUNEL X-gal luteinizing hormone minimum essential medium 4-morpholine propanesulfonic acid molecular weight outer diameter polyacrylamide gel electrophoresis phosphate-buffered saline polymerase chain reaction phenylmethylsulfonyl fluoride pregnant mares serum gonadotropin polyvinyl alcohol polyvinyl pyrrolidone sodium dodecyl sulfate standard saline citrate single stranded conformational polymorphism reverse transcription-polymerase chain reaction Tris acetate-EDTA buffer solution Tris borate-EDTA buffer solution Tris buffered saline tissue culture trophectoderm N, N, N1, N1-tetramethylethylenediamine Tdt-mediated dUTP nick-end labeling 5-bromo-4-chloro-3-indolyl-b-D-galactoside