Table Of ContentRepRoductive Medicine
a Practical guide to
Selecting gameteS
a Practical guide to
embryoS
and Selecting
Edited by
Markus Montag, PhD
iLabCoMM GmbH, International Reprolab Consulting
gameteS
St. Augustin, Germany
and
Among the many recent advances in assisted reproduction therapies (ART), improved
technologies for identifying viable oocytes, sperm, and embryos are of primary importance.
Paradoxically, the latest advances presented at conferences and symposia are often slow embryoS
to become part of the daily routine in IVF laboratories. Detailing established and developing
techniques, A Practical Guide to Selecting Gametes and Embryos provides a user-friendly
text of ready-to-use ARTs that can be utilized effectively in the lab.
In this volume, renowned embryologist and educator Markus Montag and his expert panel
highlight sophisticated and proven selection strategies and emphasize the importance of
proper lab practice in handling gametes and embryos.
topics include
• Steps undertaken for the analysis of a semen sample
• Quality control and prevention of exposure to toxins in oocyte collection
and embryo culture
• Morphological selection of gametes and embryos
• Both commonly used and innovative techniques for gamete and embryo
selection, such as oxygen respiration and time-lapse imaging
• Invasive techniques, including polar body, embryo, and blastocyst biopsies
as well as aneuploidy testing by FISH and array-CGH
Accompanied by numerous figures and descriptions, this guide to selecting gametes and Markus Montag
embryos brings the insight of international authors with knowledge and expertise, highlighting
EditEd by
practical tips and key points. The book offers a starting point for applying successful selection
strategies for reducing the rate of high-risk multiple gestations while maintaining or increasing
viable pregnancy rates.
H100479
ISBN-13: 978-1-84214-547-0
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9 781842 145470
H100479_Cover_mech.indd All Pages 4/2/14 12:29 PM
a Practical guide to
Selecting
gameteS
and
embryoS
a Practical guide to
Selecting
gameteS
and
embryoS
Edited by
Markus Montag, PhD
iLabCoMM GmbH, International Reprolab Consulting
St. Augustin, Germany
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
CRC Press
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© 2014 by Taylor & Francis Group, LLC
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Version Date: 20140224
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Contents
Preface ������������������������������������������������������������������������������������������������������������������������������������������������������������������vii
Contributors �����������������������������������������������������������������������������������������������������������������������������������������������������������ix
1. Handling Gametes and Embryos: Sperm Collection and Preparation Techniques �������������������������������1
Verena Nordhoff, Con Mallidis, and Sabine Kliesch
2. Handling Gametes and Embryos: Oocyte Collection and Embryo Culture �����������������������������������������17
Lars Johansson
3. Handling Gametes and Embryos: Quality Control for Culture Conditions �����������������������������������������39
Jason E. Swain
4. Morphological Selection of Gametes and Embryos: Sperm �������������������������������������������������������������������59
Pierre Vanderzwalmen, Magnus Bach, Olivier Gaspard, Bernard Lejeune, Anton Neyer,
Françoise Puissant, Maximilian Schuff, Astrid Stecher, Sabine Vanderzwalmen,
Barbara Wirleitner, and Nicolas H. Zech
5. Morphological Selection of Gametes and Embryos: Oocyte ������������������������������������������������������������������81
Başak Balaban and Thomas Ebner
6. Morphological Selection of Gametes and Embryos: 2PN/Zygote ����������������������������������������������������������97
Martin Greuner and Markus Montag
7. Morphological Selection of Gametes and Embryos: Embryo ���������������������������������������������������������������115
Gayle Jones and M. Cristina Magli
8. Morphological Selection of Gametes and Embryos: Blastocyst �����������������������������������������������������������131
Thomas Ebner
9. Noninvasive Techniques: Gamete Selection—Sperm ����������������������������������������������������������������������������143
Victoria Sánchez, Joachim Wistuba, and Con Mallidis
10. Noninvasive Techniques: Gamete Selection—Oocyte ���������������������������������������������������������������������������155
Laura Rienzi, Benedetta Iussig, and Filippo Maria Ubaldi
11. Noninvasive Techniques: Embryo Selection by Oxygen Respiration ���������������������������������������������������165
Alberto Tejera, Belén Aparicio, Carmela Albert, Arancha Delgado, and Marcos Meseguer
12. Noninvasive Techniques: Embryo Selection by Time-Lapse Imaging �������������������������������������������������177
Alison Campbell
13. Noninvasive Techniques: Embryo Selection by Transcriptomics, Proteomics,
and Metabolomics ��������������������������������������������������������������������������������������������������������������������������������191
Asli Uyar and Emre Seli
v
vi Contents
14. Invasive Techniques: Polar Body Biopsy ������������������������������������������������������������������������������������������������209
Markus Montag, Jana Liebenthron, and Maria Köster
15. Invasive Techniques: Embryo Biopsy at the Cleavage Stage ����������������������������������������������������������������219
Anick De Vos
16. Invasive Techniques: Blastocyst Biopsy ��������������������������������������������������������������������������������������������������233
Steve McArthur
17. Invasive Techniques: Aneuploidy Testing by FISH �������������������������������������������������������������������������������241
Semra Kahraman and Çağrı Beyazyürek
18. Invasive Techniques: Aneuploidy Testing by Array-CGH ��������������������������������������������������������������������255
Alan R. Thornhill, Christian Ottolini, Gary Harton, and Darren Griffin
19. Summary: Comprehensive Summary of Main Points by Topic �����������������������������������������������������������269
Markus Montag
Preface
The development of current technologies to identify the most viable oocyte, sperm, or embryo is one of the most
discussed topics in assisted reproduction conferences and symposia� However, most of these, sometimes fancy,
technologies are yet to be introduced in the daily routine of an in vitro fertilization laboratory, and the reasons
for this are many�
So, from the perspective of the laboratory, it would be helpful to have an overview of ready-to-use assisted
reproduction technologies, with practical guidance from those who have knowledge and expertise in the relevant
field� The resultant textbook, A Practical Guide to Selecting Gametes and Embryos, has compiled such practical
tips, accompanied by numerous figures and descriptions of methods� This book differentiates between noninva-
sive and invasive techniques; however, to my understanding, even the most sophisticated selection strategies may
fail if they are not based on a sound start� It happens frequently—that is, by looking at the most ideal endpoint,
we simply overlook the way to reach that endpoint� Practically speaking, the starting point involves good practice
in the laboratory setting in handling gametes and embryos with special focus on quality measures� This starting
point is an absolute prerequisite to be able to apply selection strategies that make sense�
I thank all the authors who have devoted their time to contribute to this illustrated textbook� They are all active
in their respective fields with numerous obligations; hence, taking on the additional task of contributing to this
book is a commitment that cannot be praised enough�
I also thank Robert Peden (Senior Editor for Reproductive Medicine, CRC Press) for his enthusiasm, support,
and continuous efforts in bringing this project to life�
Markus Montag
vii