Laboratory Manual in Assisted Reproductive Technology Kamini A Rao, MS Srinivas
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1Laboratory Manual in Assisted Reproductive Technology2
3Laboratory Manual in Assisted Reproductive Technology
Editor Kamini A Rao DGO DORCP DCh FRCOG MCh FICOG PGDMLE (Law) FNAMS President ISAR Medical Director Bangalore Assisted Conception Centre Pvt Ltd 6/7 Kumara Krupa Road High Grounds Bangalore Co-Editor MS Srinivas MSc PhD Chief Embryologist Bangalore Assisted Conception Centre Pvt Ltd Bangalore
4Published by
Jitendar P Vij
Jaypee Brothers Medical Publishers (P) Ltd
EMCA House, 23/23B Ansari Road, Daryaganj
New Delhi 110 002, India
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Laboratory Manual in Assisted Reproductive Technology
© 2006, Kamini A Rao
All rights reserved. No part of this publication should 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 written permission of the editors and the publisher.
First Edition: 2006
9788180617737
Typeset at JPBMP typesetting unit
Printed at Gopsons Paper Ltd
5to
the countless infertile couples from
whom we have learnt so much and who still inspire us to
strive harder6
7Contributors
C Mohammed Ashraf
Medical Director
CRAFT, Kodungalur
Kerala, India
Pushpa M Bhargava
Founder-Director
Centre for Cellular and Molecular Biology,
Hyderabad, Member of Expert
Group for Formulating the National Guidelines on “Accreditation” Supervision and Regulation of ART Clinics in India
Ashok K Bhattacharya
Division of Reproductive
Biology and Molecular
Endocrinology
Department of Biochemistry
University of Calcutta
Kolkata, India
Damayanti Chakraborty
Division of Reproductive
Biology and Molecular
Endocrinology
Department of Biochemistry
University of Calcutta
Kolkata, India
BN Chakravarthy
Institute of Reproductive
Medicine, Kolkata, India
Ratna Chattopadhyay
Institute of Reproductive
Medicine, Kolkata, India
Natachandra Chimote
Vasundhara Clinic and Assisted Conception Centre
Nagpur, India
Meena Chimote
Vasundhara Clinic and Assisted Conception Centre
Nagpur, India
A Radharama Devi
Diagnostic Division
Centre for DNA Finger Prints and Diagnosis, Hyderabad, India
Sulochana Gunasheela
Gunasheela IVF Centre
Bangalore, India
R Jayaganesh
Madras Andrology
Research Centre
Chennai, India
Orly Lacham-Kaplan
Centre for Early Human
Development
Monash Institute of Reproduction and Development
Melbourne, Australia
Suresh Kattera
Scientific Director, Centre for Reproductive Medicine and
Gleneagles IVF Centre
Gleneagles Hospital, Singapore
Visiting Professor, Department of Obstetrics and Gynecology and Reproductive Medicine
Sri Ramachandra Medical
College and Research
Institute, Chennai
Nalini Krishnan
Medical Director
Desai Nursing Home
Bangalore, India
PK Vijay Kumar
Bangalore Assisted Conception
Centre Pvt Ltd, Bangalore, India
TC Anand Kumar
Hope Infertility Clinic
Bangalore, India
Uday Kumar
Stem Cell Research Centre
Manipal Hospital
Bangalore, India
Vineet Malhotra ms
Muljibhai Patel Urology
Hospital, Nadiad, India
Ranjana Mangoli
Fertility Clinic and IVF Centre
Mumbai, India
Vijay Mangoli
Fertility Clinic and IVF Centre
Mumbai, India
Rajvi H Mehta
Science Director
Hope Infertility Clinic
Research Foundation, Bangalore, India
Judith Menezes
Fertility Center
Stockholm, Sweden
Amiya Mukherjee
Harley Street Fertility
Centre, London, United Kingdom
Shreyas Padgaonkar
Shreyas Infertility and IVF Centre
Mulund, Mumbai, India
Hrishikesh D Pai
Consultant Gynecologist
Medical Director
Babies and US Fertility
IVF and ICSI Centre, Lilavathi Hospital
Senior Vice President
FOGSI 2006, Mumbai, India8
Rishma Dhillon Pai
Consultant Gynecologist
Jaslok Hospital, Chairman
Food, Drugs and Medicosurgical
Equipment Committee, FOGSI, Mumbai, India
Nandita Palshetkar
Consultant Gynecologist
Lilavathi Hospital
Chairman Perinatology
Committee FOGSI
Mumbai, India
Madhuri Patil
Patil's Endoscopy and Fertility Centre
Bangalore, India
Ameet Patki
Head of the Department
ReGenesis, Centre for Assisted Reproduction, Endoscopy and Fetal Medicine
Reliance Life Sciences
Consultant Obs and Gyn, Sri
Harkisondas Hospital, Mumbai, India
K Prabhakara
Diagnostic Division
Centre for DNA Finger Prints and Diagnostics
Hyderabad, India
A Rajasekaran
Madras Andrology
Research Centre, Chennai, India
Kamini A Rao
Medical Director
Bangalore Assisted
Conception
Centre Pvt Ltd., Bangalore
India
Varsha Samson Roy
Scientific Director
CRAFT, Kodungalur Kerala and Advanced Infertility
Centre, Bangalore, India
A Henry Sathananthan
Monash Institute of Reproduction and Development
Melbourne, Australia
Shailaja Gada Saxena
Research Scientist
ReGenesis, Genetics and Embryonic Germ Stem Cell
Reliance Life Science, Mumbai, India
Kamala Selvaraj
Fertility Research Centre
GG Hospital
Chennai, India
Priya Selvaraj
Fertility Research Centre
GG Hospital, Chennai, India
Rupin Shah
Consultant Andrologist and Microsurgeon Lilavathi
Hospital and Research Centre
Mumbai, India
Peter Sjoblom
Fertility Center
Stockholm, Sweden
WD Tatna Soorya
Monash Institute of Reproduction and Development
Melbourne, Australia
MS Srinivas
Chief Embryologist
Bangalore Assisted
Conception Centre Pvt Ltd
Bangalore, India
Vasan SS
Scientific Director
Manipal ART Centre and Diacon Institute of Neurology and Andrology
Bangalore, India
Satish Totey
Stem Cell Research Centre
Manipal Hospital
Bangalore, India
A Trunson
Monash Institute of Reproduction and
Development Melbourne, Australia
Alex C Varghese
Division of Reproductive
Biochemistry and Molecular Endocrinology
Department of Biochemistry
University of Calcutta
Kolkata, India
9Preface
Over the past two decades, vast changes have occurred in virtually all aspects of infertility management creating a sense of optimism in the minds of both the treating physician and the infertile couple. At the beginning of this era concepts such as intracytoplasmic sperm injection, assisted hatching, polymerase chain reaction and preimplantation genetic diagnosis would have been considered as science fiction, whereas in actuality, they have now become common and routine techniques in the management of infertility.
Though a number of books have been brought out on the clinical aspects of assisted reproductive technology the laboratory aspects still remain a partial mystery to many. This is a book for all professionals involved in ART: Clinicians, embryologists and paramedical personnel working in an IVF/ICSI unit. This well illustrated and timely publication provides concise yet comprehensive practical information on the laboratory aspects of infertility management and at the same time attempts to bring the latest advances in laboratory techniques for infertility within reach of the practicing specialist and technician. The aim throughout is to explain issues clearly, simply, and directly in such a way that will be understood by gynecologists, infertility specialists and especially scientists and laboratory personnel. We have tried to address many questions—some scientific some purely academic—blastocyst culture, preimplantation aneuploidy screening, optimizing embryo selection and cryopreservation programs, are all questions which are still in the open. Yes or No? If yes, when and how? We do not yet have the answers!
The current aim for all those involved with ART should be to minimize the risks and complications of ART treatment at the same time optimizing results to generate treatment of high quality. We hope that this book will help in achieving this goal.
Kamini A Rao
MS Srinivas