Table Of ContentASSESSMENT OF MAMMALIAN EMBRYO QUALITY
INVASIVE AND NON-INVASIVE TECHNIQUES
ASSESSMENT OF MAMMALIAN
EMBRYO QUALITY
Invasive and non-invasive techniques
edited by
Ann Van Soom
Department of Reproduction, Obstetrics and Herd Health,
Faculty of Veterinary Medicine, Merelbeke, Belgium
and
Marleen Boerjan
Research and Development, Pas Reform Hatchery Technologies,
Zeddam, The Netherlands
Springer Science+Business Media, B.V.
A C.I.P. Catalogue record for this book is available from the Library of Congress.
ISBN 978-94-010-3909-3 ISBN 978-94-010-0343-8 (eBook)
DOI 10.1007/978-94-010-0343-8
Printed on acid-free paper
All Rights Reserved
© 2002 Springer Science+Business Media Dordrecht
Orignally published by Kluwer Academic Publishers in 2002
Softcover reprint of the hardcover 1st edition 2002
No part of this work may be reproduced, stored in a retrieval system, or transmitted
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or otherwise, without written permission from the Publisher, with the exception
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and executed on a computer system, for exclusive use by the purchaser of the work.
TABLE OF CONTENTS
List of contributors ............................................................................vii
Preface ..............................................................................................xi
Introduction Only the best conceptus will do!
But what does best mean?.........................................xiii
Martin H. Johnson
Chapter 1: Human embryo morphology and developmental
capacity.................................................................................1
Mina Alikani, Sasha Sadowy and Jacques Cohen
1. Introduction...................................................................................1
2. EggCyte™ Embryology Database.................................................1
3. Zygote Morphology.......................................................................2
4. Cleavage Stage embryos................................................................7
5. Morulae and Blastocysts..............................................................18
6. Conclusions and future directions...............................................24
Chapter 2: Morphological evaluation of embryos in domestic
species.................................................................................31
Sybrand Merton
1. Introduction.................................................................................31
2. Developmental processes in embryos..........................................33
3. Classification of embryos by stage of development....................38
4. Classification of embryos by quality grade.................................40
5. Relationship between embryo morphology evaluation and
pregnancy rates............................................................................43
6. Factors affecting embryo evaluation...........................................45
7. Embryo morphology in other domestic species...........................48
8. Conclusion...................................................................................49
Chapter 3: Metabolic markers of embryo viability...........................55
Isabelle Donnay
1. Introduction.................................................................................55
2. General characteristics of embryo metabolism............................56
ii
3. Non-invasive techniques of evaluation........................................62
4. Evaluation of embryo viability after embryo manipulation........78
5. Conclusions.................................................................................80
Chapter 4: Embryo quality and freezing tolerance:
cryopreservation of human embryos...............................93
Josiane Van der Elst
1. Introduction.................................................................................93
2. Selection of embryos for cryopreservation: features related to
freezing tolerance........................................................................95
3. Conclusion.................................................................................110
Chapter 5: Factors influencing cryopreservation of domestic
animal embryos................................................................121
Alban Massip and S. P. Leibo
1. Introduction...............................................................................121
2. Factors affecting freezing tolerance...........................................124
3. Cryobiological aspects...............................................................129
4. Possible methods to improve freezing resistance......................131
5. Conclusion.................................................................................133
Chapter 6: Timing of embryo development......................................139
Barry Bavister
1. Introduction...............................................................................139
2. Timing of embryo development as an indicator of viability.....140
3. Genetic regulation of embryo development timing...................144
4. Timing of events during fertilization.........................................145
5. Culture media effects on timing of embryo development.........146
6. Timing of human embryo development....................................148
7. Summary and conclusions.........................................................151
Chapter 7: Biopsy and preimplantation diagnosis of human
embryos.............................................................................157
Deborah Taylor and Maria Tachataki
1. Introduction...............................................................................157
2. Preimplantation genetic diagnosis (PGD).................................158
3. Obtaining oocytes and embryos................................................158
4. Methods of oocyte and embryo biopsy......................................159
iii
5. Methods of diagnosis.................................................................163
6. Use of PGD technology for evaluation of mammalian
embryo quality...........................................................................171
Chapter 8: Biopsying and genotyping cattle embryos.....................178
Erik Mullaart
1. Introduction...............................................................................178
2. Embryo biopsy methods............................................................179
3. Pregnancy rates..........................................................................182
4. DNA tests performed on the biopsy..........................................185
5. Conclusion.................................................................................191
Chapter 9: Multiphoton microscopy for imaging mammalian
embryos.............................................................................195
Jayne M. Squirrell
1. Introduction...............................................................................195
2. Multiphoton microscopy basics.................................................196
3. Embryo culture for imaging......................................................199
4. Viability (laboratory species)....................................................201
5. Imaging depth (domestic species).............................................205
6. Maximizing information from rare specimens
(non-human primate).................................................................207
7. Potential future directions..........................................................209
8. Conclusion.................................................................................211
Chapter 10: Is the frequency of chromosome abnormalities
influenced by in vitro techniques?..................................219
Dorthe Viuff, Torben Greve and Preben Dybdahl Thomsen
1. Introduction...............................................................................219
2. Methodology..............................................................................221
3. Chromosome abnormalities in embryos....................................225
4. Is the incidence of chromosomal abnormalities increased by
in vitro culture?..........................................................................230
5. Conclusion.................................................................................232
iv
Chapter 11: Differential staining of inner and outer cells
to assess mammalian embryo quality.............................237
Ann Van Soom, Marleen Boerjan and Kate Hardy
1. Introduction...............................................................................237
2. Embryonic development............................................................238
3. Differential staining of the blastocyst........................................242
4. Conclusion.................................................................................259
Chapter 12: Apoptosis in mammalian embryos.................................267
Kate Hardy and Sophie Spanos
1. Introduction...............................................................................267
2. Definitions of cell death............................................................270
3. Characteristic features of apoptosis in mammalian embryos:
incidence and detection.............................................................271
4. Examples of application............................................................284
5. Advantages and disadvantages of different techniques.............286
6. Conclusions...............................................................................288
Chapter 13: Electron microscopy of pre-hatching mammalian
embryos: the ultrastructure of cattle, swine and
horse embryos..................................................................295
Poul Maddox-Hyttel1 and Marleen Boerjan
1. Introduction...............................................................................295
2. Basic considerations on the ultrastructure of pre-hatching
mammalian embryos..................................................................296
3. Ultrastructure of the oocyte, zygote and pre-hatching
embryo in cattle.........................................................................300
4. Ultrastructure of the zygote and pre-hatching embryo in swine328
5. Ultrastructure of the zygote and pre-hatching embryo in horse334
Chapter 14: Differences in gene expression patterns related to
origin of preimplantation embryos................................341
Christine Wrenzycki and Heiner Niemann
1. Introduction...............................................................................341
2. Methodological aspects of mRNA expression analysis.............343
3. Gene expression patterns in bovine preimplantation embryos..346
4. Alterations in gene expression patterns of preimplantation
rodent embryos..........................................................................357
v
5. Gene expression patterns in human embryos............................362
6. Summary and concluding remarks............................................363
Chapter 15: Protocols
Appendix I: The electron microscope............................................371
Poul Maddox-Hyttel
The transmission and scanning electron microscope.................371
Microscopical resolution...........................................................372
Specimen preparation for transmission electron microscopy....374
Appendix II: Processing of eggs for transmission electron
microscopy..................................................................376
Poul Maddox-Hyttel
Appendix III: Human preimplantation diagnosis pcr
and RT-PCR...............................................................379
Deborah Taylor
Appendix IV: Human preimplantation diagnosis fluorescence
in situ-hybridization (FISH) on individual
blastomeres.................................................................382
Deborah Taylor
Appendix V: Fluorescence in situ-hybridization (FISH) on
metaphases in bovine embryos..................................384
Dorthe Viuff
Appendix VI: Differential staining....................................................388
Ann Van Soom
Index ......................................................................................392
Colour plates ......................................................................................401
LIST OF CONTRIBUTORS
Alikani M
Institute of Reproductive Medicine and Science, Saint Barnabas
Medical Center, 101 Old Short Hills road, Suite 501, West
Orange, NJ 07052, USA
Bavister BD
Audubon Institute, Center for Research of Endangered Species,
University of New Orléans, 14001 River Road, New Orleans, LA
70131, USA
Boerjan ML
PasReform, Hatchery technologies, PO Box 2, NL-7038 ZG
Zeddam, the Netherlands
Cohen J
Institute of Reproductive Medicine and Science, Saint Barnabas
Medical Center, 101 Old Short Hills road, Suite 501, West
Orange, NJ 07052, USA
Donnay I
Université Catholique de Louvain, Unité des Sciences Vétéri-
naires, Place Croix du Sud 3, B-1348 Louvain-la-Neuve,
Belgium
Greve T
Department of Clinical studies, Reproduction, The Royal
Veterinary and Agricultural University, Dyrlægevej 68, 1870
Frederiksberg C, Copenhagen, Denmark
Hardy K
Department for Reproductive Science and Medicine, Institute of
Reproductive and Developmental Biology, Imperial college,
Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK
Johnson MH
Department of Anatomy, University of Cambridge, Downing
Street, Cambridge CB2 3DY, UK
viii
Leibo SP
Audubon Institute, Center for Research of Endangered Species,
University of New Orléans, 14001 River Road, New Orléans, LA
70131, USA
Maddox-Hyttel P
Department of Anatomy and Physiology, The Royal Veterinary
and Agricultural University, Groennegaardsvej 7, 1870 Frede-
riksberg C, Copenhagen, Denmark
Massip A
Université Catholique de Louvain, Unité des Sciences
Vétérinaires, Place Croix du Sud 3, B-1348 Louvain-la-Neuve,
Belgium
Merton S
Holland Genetics, ET-IVP Laboratory, Broekstraat 5b, 7217 SL
Harfsen, the Netherlands
Mullaart E
Holland Genetics, PO Box 5073, 6802 Arnhem, the Netherlands
Niemann H
Department of Biotechnology, Institute of Animal Science,
Federal Agricultural Research Center (FAL), Höltystr. 10, D-
31535 Neustadt, Germany
Sadowy S
Institute of Reproductive Medicine and Science, Saint Barnabas
Medical Center, 101 Old Short Hills road, Suite 501, West
Orange, NJ 07052, USA
Spanos S
Department for Reproductive Science and Medicine, Institute of
Reproductive and Developmental biology, Imperial college,
Hammersmith Hospital, Du Cane Road, London, W12 ONN, UK
Squirrell JM
Laboratory of Moleuclar Biology, University of Wisconsin, 1525
Linden Dr., Madison, WI 53717, USA
Tachataki M
Institute of Reproductive Biology, Faculty of Veterinary
