Male Infertility: Sperm Diagnosis, Management and Delivery Jayant G. Mehta, Bryan Woodward
INDEX
Note: Page numbers in bold or italic refer to tables or figures respectively.
A
Accessory sex glands 12
Acridine orange test (AOT) 48, 49
Agglutination 12, 66
Aggregation 66
Agitator 10, 10
Alcohol, effects of, on fertility 104
Alpha-receptor blocking agents, and retrograde ejaculation 90
Androgens 4
Aneuploidy testing 115
Annexin-V 113
Antibiotics, in culture media 75, 75
Antidepressants, and retrograde ejaculation 90
Antifertility factor (AF-1) 38
Antioxidant therapy 58, 59
Antisperm antibodies (ASAs) 12, 17, 45, 51, 5960
bypassing of, with ART 5354
epidemiology of 51
formation
causes of 53
prevention of 53
immunological infertility and 5152
infertility 51
treatment of 5354
testing for 51, 52
vasectomy and 53, 53
Array comparative genomic hybridization (array-CGH) technology 115
ART see Assisted reproduction therapy (ART)
ASAs see Antisperm antibodies (ASAs)
Aspermia 69
Assisted reproduction therapy (ART) 37, 40, 45, 4849, 53, 7179, 93, 97
culture media for 2527 (see also Culture media)
impact of, on sperm function tests 49 (see also Sperm function tests (SFTs))
sperm morphology in, influence of 2122
Atomic force microscopy 114
Azoospermia 5, 13, 65, 67, 81, 93, 98
non-obstructive 68, 69
obstructive 68, 69, 81
B
Bacteriospermia 76
Blood-borne virus (BBV), screening for 72
Bulbourethral gland 1
C
Cancer patients
sperm banking for 98100
sperm cryostorage, legal issues related to 122
Candida albicans 77
Capacitation 28, 37 see also Sperm preparation
Cellometer counting chamber 12, 13
Cell-Vu 12, 13
Cervical mucus 47
Chlamydia trachomatis 58
Chromosome microdeletions 48
Clomiphene (clomiphene citrate) 62, 69, 105
Coitus interruptus 8, 66
Collection artifacts 66
COMET (Single cell gel electrophoresis) assay 48, 49
Competence matrix 119, 119
Computer-aided sperm analyses (CASA) 17
Computer-controlled sperm freezing 95, 96
Congenital bilateral absence of vas deferens (CBAVDs) 69
Congenital rubella syndrome 121
Corpora cavernosa 2
Corpus spongiosum 2
CryoBioSystem
high-security straws 94, 95
high-security tubes/vials 95, 95 Cryoprotectant medium (CPM) 94
Cryptorchidism 57, 57
Culture media 2526, 26, 26
amino acids in 3132
antibiotics in 75, 75
back-to-nature principle 25, 26
development of 27, 27
ejaculated sperm journey and 28, 28
ionic composition 29
pH 29, 2931, 30, 31
phosphate 29
let-the-embryo-choose principle 25
macromolecules in 32
nutrients in 28
glucose 29
lactate 2829
pyruvate 28
osmolality of 32, 3233
preimplantation development and 2728
sequential and monoculture media, constituents of 28
sperm selection process and 25, 25
vitamins in 32
Cytomegalovirus (CMV) 7778
D
Diff-Quik stain 15
Dimethylsulphoxide (DMSO) 94
Direct swim-up technique 42 see also Sperm preparation
Discontinuous Percoll gradient method 3839
Dithiothreitol (DTT) 127
Donor sperm 93
Donor sperm banking 123
E
Earle's balanced salt solution 27
Efferent ductules 2
Egg yolk, as extender 94
Ejaculation
antegrade 90
normal 89, 89
retrograde (see Retrograde ejaculation (RE))
Ejaculatory duct 2
Ejaculatory duct obstruction (EDO) 65, 69
Electroejaculation 69
Enzyme-linked immunosorbent assay (ELISA) 52
Eosin–nigrosin test 16
Epididymal obstruction 69
Epididymis 1, 2, 3
Escherichia coli (E. Coli) 75
Ethical issues, related to ART 123124
European Tissues and Cells Directive (EUTCD) 7174, 8485, 94, 120, 121
European Union (EU) Directive, for sperm laboratories 119
authorities and enforcement 119120
cryopreservation 121
documentation 120121
equipment and materials 120
facilities 120
laboratory testing 121
personnel 119, 119
quality management 121
traceability 120
EUTCD see European Tissues and Cells Directive (EUTCD)
Evie insemination device 104, 105
Extenders 94
External quality assessment (EQA) service 17
F
Flow cytometry 46, 52
Fluorescence-activated flow cytometry 126
Fluorescent in situ hybridization (FISH) 115, 126127
Follicle-stimulating hormone (FSH) 4, 83, 105
G
Gardnerella vaginalis 58
Genital tract infections 69
Globozoospermia 15
Glucose, in culture media 29
Gonadotropin-releasing hormone (GnRH) 4, 5
Gonadotropins 105
H
Ham's F10 or T6 solution 27
Hematospermia 11
Hemizona assay 48, 48
Hemocytometer chamber 1213, 14
Hepatitis B virus (HBV) 71
Hepatitis C virus (HCV) 71, 74
Hoechst 33342 126
human chorionic gonadotropin (hCG) 67, 105, 106, 108
Human immunodeficiency virus (HIV) 71, 74
Human serum albumin 30, 31
Human sperm cryobanking 93
indications for 93
autoconservation 93
donor sperm conservation 93
methods of 9398
cooling/freezing method 9596
cryoprotectants 94
packaging container 9495, 95
pre-freeze and post-thaw preparations 9798
sperm freezing 9394
sperm thawing 97
storage tanks and facilities 97, 97
storage temperature 9697
oncological patients and 98100
Human sperm toxicity test (HSTT) 45, 49
Human tubal fluid (HTF) 27
Hyaluronan-binding assay 48
Hyaluronic acid-based sperm binding assays 113, 113
4-(2-Hydroethyl)-1-piperazineethanesulphonic acid (HEPES) 30
Hypogonadotropic hypogonadism (HH) 5, 67
Hypo-osmotic swelling (HOS) test 16, 27, 46, 46, 84, 112
Hypospadias 7, 57, 57
I
ICSI see Intracytoplasmic sperm injection (ICSI)
Immotile cilia syndrome 5
Immotile sperm viability assay 112
Immunobead test (IBT) 52
Infertility, male 5, 7, 51 see also Semen analysis (SA)
drugs for treatment of 6162
retrograde ejaculation and 89
substances causing 5758, 59
varicocele and 60 (see also Varicocele)
Insemination program 103
emotional support and counseling for 109
fertility nurse's role in 103
intracervical insemination 104, 105
intrauterine insemination 104, 105
loading of IUI catheter 107
ovulation induction and cycle monitoring 104105, 106
patient preparation for insemination 107
planning of 104, 104
semen sample collection 106107
standard operating procedures (SOPs) for 104
steps after insemination procedure 108
ultrasound scanning 105106
vaginal and pelvic examinations and insemination process 107108
Internal quality control (IQC) 17
Intracytoplasmic morphologically selected spermatozoa injection (IMSI) 111, 112
Intracytoplasmic sperm injection (ICSI) 53, 54, 60, 77, 8182, 91, 93, 111 see also Surgical sperm retrieval (SSR)
and bacterial contamination 77
risk of infection during 77
Intrauterine insemination (IUI) 22, 41, 53, 54, 67, 69, 7275, 9799, 104106, 108, 126
In vitro fertilization (IVF) 38, 4041, 45, 49, 53, 54, 71, 7478, 97, 111
pH-ranges of sequential and monoculture media for 30, 31
sperm morphology in, influence of 21
IUI see Intrauterine insemination (IUI)
K
Kremer test 47
Kurzrok–Miller test 47
L
Lactate, in culture media 2829
Laser tests, for testing viability in immotile sperm 112, 113
Leukocytospermia 58
Leydig cells 4
Liverpool Solution 91, 91
Luteinizing hormone (LH) 4, 105
M
Makler counting chamber 12, 13
Male sex organs
anatomy of 1, 12, 2
accessory sex glands 12
epididymis 1
penis 2
scrotum 2
testes 1, 2
blood supply of 23
innervation of 3
Microbial contamination 7576
laboratory practice for reducing 7677
Microepididymal sperm aspiration (MESA) 81 see also Surgical sperm retrieval (SSR)
Microscopic sperm organelle morphological evaluation (MSOME) 111, 112
Microsort see Fluorescence-activated flow cytometry
Mixed agglutination reaction (MAR) 52, 60, 66
3-(N-morpholino)-1-propanesulfonic acid (MOPS) 30
Motility, sperm, assessment of 1112
Multitube direct swim-up technique 42 see also Sperm preparation
Mycoplasma genitalium 77
N
Necrozoospermia 16
Neubauer hemocytometer 12, 14
Nurse-led clinics 103 see also Insemination program
O
Oligozoospermia 66, 70, 98
Oocyte–cumulus–corona complex (OCC) 39
Ovarian hyperstimulation syndrome (OHSS) 104
Oxidative stress induced male fertilit 58, 59
P
Pap-stain 15
‘Partner donation’ 7172
Penis 2, 3
Pentose phosphate pathway (PPP) 29
Pentoxifylline 112
Percoll 38, 41
Percutaneous sperm aspiration (PESA) 81 see also Surgical sperm retrieval (SSR)
pH buffering 3031
Phenol red, use of, as a pH indicator 31
pH, semen 11
Phthalates 57
Plasmalogens 37
PLCzeta 114
Polyzoospermia 66
Postmortem sperm retrieval (PMSR) 123
Potassium simplex optimized medium (KSOM) 27
Primordial germ cells 3
Prostaglandins, and uterine contractions 38
Prostate glands 12
Pyruvate, in culture media 28
Q
Quinacrine chloride 125126
R
Raman spectroscopy 114
Reactive oxygen species (ROS) 58
Rete testes obstruction 69
Rete testis 1
Retrograde ejaculation (RE) 69, 89, 89
causes of 89
bladder neck and prostate surgery 8990
congenital 90
diabetes mellitus 90
drugs 90
operative nerve damage 90
spinal injury 90
treatment of 90
bladder conditions, adjustments of 91, 91
drug treatments 90
operative measures 90
retrieving sperm from urine 91
special maneuvers 90
Rotator 10, 10
Round cells 1617
S
Scrotum 2, 3
Semen, abnormal 65
aggregation and agglutination 66
color 66
fructose 65
low volume 65
motility 67, 69
pH 65
round cells 67, 68
sperm concentration 6667, 67, 68
viscosity and liquefaction 66
zinc and neutral α-glucosidase 6566
Semen analysis (SA) 7, 45, 6567 see also Semen, abnormal
computer-aided 17
laboratory area for 10
macroscopic assessment
color 11
liquefaction and viscosity 11
pH 11
volume 1011
microscopic assessment 11
antisperm antibodies 17
count 1213, 13, 14
equipment for 11
morphology 13, 15, 1516, 16
motility 1112, 12
round cells 1617
vitality 16
performers of 7
production to, time from 10, 10
quality assurance, need for 17
reasons for 7
report of 1718, 18
sample production for
home production 8
patient information and instructions 810
production room 78, 66
specimen container 8
staff training for 7
and treatment
aspermia 69
azoospermia 6769
genital tract infections 69
low/high volume semen 67
sexual dysfunction 6970
Semen quality, poor
environment and 57, 5758, 58
immunological causes of 5960
inflammatory and infectious causes of 58, 59, 59
varicocele and 6061, 61
Seminal plasma 37
factors in, affecting fertilization 3738
removal of 3738
and sperm motility and viability 37
sperm separation from 3839
density gradient centrifugation 3839
swim-up technique 38
Seminiferous tubules 1, 2, 2
Sertoli cells 4
Sex selection 125
Microsort for 126128
description of 126127, 127
efficacy of sorting process 127, 127
history and background 126
outcomes 127128, 128
natural methods for
diet and vaginal pH 125
timing of intercourse 125
X- and Y-bearing sperm, physical separation of 125126
Sildenafil citrate (Viagra) 69
Specimen container, semen 8
Spermatogenesis
androgen regulation of 45
hormonal dependency of 5
organization of 34, 4
Spermatogonia 3
Spermatozoa 1, 2
function of 5
production of 3, 3, 4
Sperm bioassay 4849
Sperm chromatin dispersion test (SCD) 48, 49
Sperm chromatin structure assay (SCSA) 48, 49
Sperm count 12
Sperm cryostorage, legal issues related to 121122
cancer patients and 122
information to patients 122
posthumous-assisted reproduction 122123
postmortem sperm retrieval 123
storage contracts for patients 122
Sperm DNA fragmentation index 48, 49
Sperm DNA, integrity of 113114
Sperm function tests (SFTs) 45
hypo-osmotic swelling test 46, 46
objective of 45
sperm acrosome reaction 46
sperm–cervical mucus interaction 4647
sperm chromatin assessment 48
sperm longevity assessment 46
sperm maturity appraisal 48
sperm–zona pellucida binding 4748
stress tolerance test 45
vitality testing 46
Sperm head polarization microscopy 114
Spermiation 3
Spermiogenesis 3, 3
Sperm morphology
analysis of 13, 15, 1516, 16
patterns
G-pattern 2122
N-pattern 2122
P-pattern 2122
reference values 2122
role of, in assisted reproduction 2122
WHO approach on semen thresholds 22
Sperm-omics 115
Sperm–oocyte activation process 114
Sperm preparation 37, 74, 74
principle of 39, 3940
protocol for harvesting motile sperm 40
density gradient sperm separation protocol 41, 4142
direct swim-up technique 42
multitube direct swim-up technique 42
semen collection 40
swim-up protocol 4041
sperm separation from seminal plasma in 3738
density gradient centrifugation 3839
swim-up technique 38
Sperm-rise test 38
Sperm selection technologies 111, 111115
Sperm washing 38, 7475, 97
Sperm zona binding tests (ZBT) 4748, 48
Split pot procedure 53
SSR see Surgical sperm retrieval (SSR)
Stress tolerance test 45
Surgical sperm retrieval (SSR) 13, 67, 8182
considerations in
digestive enzyme, use of 84
DNA damage, levels of 83
fertilization rates and pregnancy outcome 83
fresh or frozen sperm 83
likelihood of obtaining sperm 8283
motility enhancement 84
single or multiple biopsies 83
equipment for 85
European regulatory requirements on 8485
fresh and post-thaw preparation 8687
ICSI procedure 87, 87
medium for 85
preparation for 85
samples, cryopreservation of 86
techniques
microepididymal sperm aspiration 81
percutaneous sperm aspiration 81, 85
testicular sperm aspiration 81, 81
testicular sperm extraction 81, 82, 8586
thawing in 86
T
Tadalafil (Cialis) 69
Testes 1, 2
blood supply to 23
endocrine signals, response to 4
germ cells in, development of 3
nerve supply to 3
spermatozoa, production of 3, 3, 4 (see also Spermatogenesis)
Testicular cancer 57, 58
Testicular dysgenesis syndrome 57
Testicular sperm aspiration 81, 81 see also Surgical sperm retrieval (SSR)
Testicular sperm extraction (TESE) 53, 81, 82 see also Surgical sperm retrieval (SSR)
Testicular stem cells, freezing of 99100
Testosterone 1, 4
Theophylline 112
Tight foreskin 7
Tray agglutination test 52
TUNEL (Terminal deoxynucleotidyl transferase dUTP nick and labeling) assay 48, 49
U
Unstimulated intrauterine insemination (IUI) 103, 104 see also Insemination program
Ureaplasma urealyticum 58
Urethra 2
V
Vardenafil (Levitra) 69
Varicocele 3, 7, 60, 61
conclusions and recommendations for 60
and poor semen quality 60
repair of, method of 6061, 61
Vas deferens 1, 2
Vasectomy 51, 53, 69, 81
Viral infections, screening for 71
Vitality test 46
Vitrification 95, 99100
Volume, semen 1011
W
Wet split procedure 53
World Health Organization (WHO) 6, 1013, 16, 17, 21, 22, 46, 65, 69
X
Xenotransplantation 99
Y
Y chromosome microdeletion testing 83
Z
Zinner syndrome 11
Zona pellucida (ZP) 5
×
Chapter Notes

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1MALE INFERTILITY SPERM DIAGNOSIS, MANAGEMENT AND DELIVERY2
3MALE INFERTILITY SPERM DIAGNOSIS, MANAGEMENT AND DELIVERY
Jayant G. Mehta, PhD, DipRCPath Director of Sub-Fertility Laboratory and Quality Control Manger Barking, Havering and Redbridge University Hospitals NHS Trust Queen's Hospital Romford Essex, UK Bryan Woodward, PhD, FRCPath Consultant Embryologist and Clinical Scientist IVF Consultancy Services Leicester, UK
4© 2014 JP Medical Ltd.
Published by JP Medical Ltd, 83 Victoria Street, London, SW1H 0HW, UK
Tel: +44 (0)20 3170 8910
Fax: +44 (0)20 3008 6180
The rights of Jayant G. Mehta and Bryan Woodward to be identified as editors of this work have been asserted by them in accordance with the Copyright, Designs and Patents Act 1988. All rights reserved. No part of this publication may be reproduced, stored or transmitted in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, except as permitted by the UK Copyright, Designs and Patents Act 1988, without the prior permission in writing of the publishers. Permissions may be sought directly from JP Medical Ltd at the address printed above.
All brand names and product names used in this book are trade names, service marks, trademarks or registered trademarks of their respective owners. The publisher is not associated with any product or vendor mentioned in this book.
Medical knowledge and practice change constantly. This book is designed to provide accurate, authoritative information about the subject matter in question. However readers are advised to check the most current information available on procedures included and check information from the manufacturer of each product to be administered, to verify the recommended dose, formula, method and duration of administration, adverse effects and contraindications. It is the responsibility of the practitioner to take all appropriate safety precautions. Neither the publisher nor the editors assume any liability for any injury and/or damage to persons or property arising from or related to use of material in this book.
This book is sold on the understanding that the publisher is not engaged in providing professional medical services. If such advice or services are required, the services of a competent medical professional should be sought.
Every effort has been made where necessary to contact holders of copyright to obtain permission to reproduce copyright material. If any have been inadvertently overlooked, the publisher will be pleased to make the necessary arrangements at the first opportunity.
9781907816468
British Library Cataloguing in Publication Data
A catalogue record for this book is available from the British Library
Library of Congress Cataloging in Publication Data
A catalog record for this book is available from the Library of Congress
JP Medical Ltd is a subsidiary of Jaypee Brothers Medical Publishers (P) Ltd, New Delhi, India.
Publisher:
Richard Furn
Development Editor:
Gavin Smith
Design:
Designers Collective Ltd
Typeset, indexed, printed and bound in India.
5Foreword
The editors, both authorities in this field, have gathered a group of active, eminent scientists to produce a book that addresses the practical needs of trainees, small community hospitals and nurse-led programmes in the UK, the rest of Europe and globally.
The contributing authors have very much focussed on providing information in plain language, encouraging the novice to read more, as well as facilitating the experienced practitioner in updating their expertise. The use of line drawings reinforces the crisp text. The order of the contributions is logical, introducing the reader to this poorly-understood area of the infertility equation.
Day-to-day challenges for patients as well as practitioners are addressed by providing tricks and tips to indicate clearly the recommended patient pathway. These can be adapted to a wide range of patient expectations, cultural influences and individual circumstances. The inclusion of a sound protocol for ‘home production’ is notable and will lead not only to stress avoidance for patients and the andrology team, but also to a lesser demand for emergency strategies such as oocyte freezing or vitrification and surgical sperm retrieval.
Up-to-date reference values are highlighted in an early chapter as are the benefits of using the most advanced consumables available. Authoritative reviews of cutting-edge matters such as sperm DNA fragmentation tests and their role are welcome, as well as clarification of the relevance of immunological phenomena in modern infertility practices.
How the needs of our changing society and its behaviour are met with state-of-the-art ART practices are reflected in the valuable section on virology, surgical sperm retrieval and cryobiology, as well as the recognition of the value of nurse-led services.
This excellent volume is rounded off by a useful look to the future, the legal consequences of clinical services and gender choice issues, as well as quality management systems.
Male Infertility should be essential reading for all trainees and have a place on the shelves of all centres, large and small, that provide infertility services.
Gedis Grudzinskas
MBBS BSc MD FRANZCOG FRCOG
Consultant in Infertility and Gynaecology
Harley Street
London, UK
6
7Preface
Male infertility is emerging as a major factor affecting natural conception. Assisted reproductive technology (ART) has helped couples with male infertility to achieve a pregnancy, but only after a careful analysis of the sperm physiology. In this book, fourteen renowned scientists review the study of male infertility and treatment options using the tools and techniques currently available in the andrology laboratory, translating the information into clinical diagnosis and treatments.
The significance of technological advances in the treatment of male infertility is highlighted, including sperm preparation for insemination, the importance of cryopreservation, techniques for obtaining sperm after retrograde ejaculation and the preparation of sperm from men who are infected with blood-borne viruses.
The book begins with an introduction to the anatomy and physiology of the male and spermatogenesis. The importance of a standardized WHO-recommended semen analysis is the basis of the second chapter, which describes various diagnostic assessments that allow the reader to better understand the importance of sperm parameters and use this information to recommend the most appropriate treatment. The significance of using the correct medium with optimum constituents to maintain sperm viability and optimize the physiological pH is covered in the fourth chapter. Different sperm preparation methods are described in the fifth chapter.
Whilst a basic semen analysis provides basic information, the sixth chapter addresses further diagnostic tests that might help to establish the functionality of sperm from a semen sample. The seventh chapter deals with the immunological contributions to male infertility and how to overcome them and the eighth chapter covers the causes and effects of poor semen quality. While it is generally accepted that treatment options are similar worldwide, the ninth chapter describes subtle differences in practice. The importance of the cryopreservation of sperm is described in detail in the thirteenth chapter and highlights the existence of physiological similarities and differences between fresh and frozen semen samples. The role of the fertility nurse in a clinical setting offering intrauterine insemination (IUI) is described in great detail in the fourteenth chapter, allowing practicing nurses to perfect their techniques. The remaining chapters deal with retrograde sperm preparation, microsurgical aspiration techniques, preparation of semen from a serologically-positive patient, sperm and legal aspects, and sex selection of sperm. Finally, since no treatment should be permitted without informed consent, the appendix provides the reader with links to download the necessary forms from the website of the Human Fertilisation and Embryology Authority (HFEA), a regulatory body responsible for licensing all of the ART clinics in the UK.
Some overlap of subjects may be detected in different chapters, but this has been permitted in order to allow different viewpoints to be provided for the reader. However, we have made every effort to avoid repetition. Readers may also notice occasional conflicts of opinion between different authors, and the opinions expressed do not necessarily reflect the views of the editors. As editors, it is our duty to allow our authors a free rein!
This book serves as a guide to evidence-based clinical practice and as a reference text for those embarking on their andrology quest, be they medical students, embryologists, biomedical scientists, nurses or sub-specialty trainees in ART.
Jayant G. Mehta
Bryan Woodward
March 2014
8
9Acknowledgements
The success and final outcome of this book required much guidance and assistance from many people. Whatever I have done is only as a result of such help and I thank them all here, even if not specifically by name.
First and foremost, I am extremely grateful to my co-editor Dr Bryan Woodward, who came up with the idea of the book and supported me as an excellent co-editor with his patience, understanding and encouragement throughout.
The contribution of excellent review articles from all the contributors, some friends, some my peers and all eminent scientists in their own field have helped to shape this book. I sincerely thank each and every one for agreeing to contribute their knowledge and hard work.
I would also like to extend my appreciation and thanks to Geoff Greenwood, Gavin Smith and Richard Furn at JP Medical Ltd for their continuous support and encouragement.
Since many of the chapters required the use of images and figures whose copyrights were held by others, both Bryan and I are grateful to all copyright holders for granting us permission to make use of their material.
I would also like to pay tribute to the teachings of my religious ‘Guru’ – ‘Sai Baba’ of shirdi, for his guidance and inspiration in ensuring that this book was completed.
Finally, I am extremely indebted to one special individual who has always stood by my side and whose support, encouragement, tolerance and understanding have made the realization of this book possible – my wife, Minaxi Mehta.
JGM
I would like to express sincere thanks to my co-editor Dr Jayant Mehta, and echo the gratitude to all of our contributors, and to all of the staff at JP Medical Ltd. Thank you for your patience! I am also indebted to friends and family – there are too many to mention, but you know who you are!
BW
10Contributors