Lyn Healy Ludmila Ruban Atlas of Human Pluripotent Stem Cells in Culture 123 Atlas of Human Pluripotent Stem Cells in Culture Lyn Healy (cid:129) Ludmila Ruban Atlas of Human Pluripotent Stem Cells in Culture Lyn Healy Ludmila Ruban United Kingdom Stem Cell Bank Department of Biochemical Engineering National Institute for Biological Standards Faculty of Engineering Science and Control University College London Potters Bar London Hertfordshire UK UK ISBN 978-1-4899-7506-5 ISBN 978-1-4899-7507-2 (eBook) DOI 10.1007/978-1-4899-7507-2 Springer New York Heidelberg Dordrecht London Library of Congress Control Number: 2014953597 © Springer Science+Business Media New York 2015 T his work is subject to copyright. 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Violations are liable to prosecution under the respective Copyright Law. T he use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. While the advice and information in this book are believed to be true and accurate at the date of publication, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Printed on acid-free paper Springer is part of Springer Science+Business Media (www.springer.com) In loving memory of Professor Donald S. Munro, who taught me tissue culture and always believed in me. Ludmila Ruban This book is dedicated to my Mum and Jude, who have supported me always, and it is in memory of my Dad, who was the reason that I went into science. Lyn Healy Preface by Lyn Healy T he derivation of human embryonic stem cells was described in “Embryonic stem cell lines derived from human blastocysts” by James A. Thomson et al. in 1998. Shortly after this dis- covery, work on this cell type in the U.K. commenced in the laboratories of Professor Peter Andrews and Professor Harry Moore in the University of Sheffi eld. Indeed, by 2003, the groups in Sheffi eld had derived their own lines and were running training courses in the growth and characterization of human embryonic stem cell lines. I t was during the preparation for one of these courses that Ludmila and I fi rst met. We both had vast experience in cell culture, and Ludmila had the good fortune to have been involved in both the derivation and extended culture of these lines and their complex characterization. I had come into the area from a haemopoietic stem cell culture background with some experi- ence in mouse embryonic stem cell culture, and I was eager to broaden my skill set to include human embryonic stem cells. I had eight weeks to be trained so that I could help with the practical training course. Ludmila was keen to train me, and among the fi rst things that she did was to introduce me to her extensive collection of printed images of human embryonic stem cells displaying various levels of the differentiated state. With the aid of these images, she carefully explained to me what I should look for in the colonies in order to make a morphologi- cal judgment on their status. I took this learning back with me to the U.K. Stem Cell Bank when we were starting our banking processes, and many of our staff have passed through the portals of the University of Sheffi eld Laboratories and have received training from Ludmila. When Ludmila moved to University College London in 2006, we were able to work more closely, and she suggested that we put together an atlas. We both believe that a picture really is “worth a thousand words.” It is still the case that human pluripotent stem cells, including the human ES-like induced plu- ripotent stem cells, are generally assessed during their growth and expansion purely on mor- phology. This atlas is intended as a laboratory reference manual for scientists new to the fi eld as well as for those with more experience, and we hope that it will be of value to those at the bench who rely on morphology as the basis for judging, subjectively and in isolation, the qual- ity of pluripotent stem cells in culture. We hope that this atlas will prove to be a valuable resource in the laboratory for researchers at all levels of experience. Potters Bar, Hertfordshire, UK Lyn Healy vii Preface by L udmila Ruban Saying that I am not well known would be an overstatement. I am not known in the exquisite world of stem cell research. Nevertheless, in 2001, I was fortunate to join Peter Andrews’ and Harry Moore’s laboratory in Sheffi eld University, which later became the Centre for Stem Cell Biology. We were one of the fi rst in the U.K. and Europe to start working with the Wisconsin human embryonic stem cell lines. The Centre was awarded one of the fi rst two licenses granted by the U.K. Human Fertilisation and Embryology Authority (HFEA) for derivation of human embryonic stem cells. I was also actively involved in the training of national and international researchers within the Centre and coordinated, organized, and taught at the Centre for Academics’ annual practical training course, “Working with Human Embryonic Stem Cells.” I met Lyn eleven years ago when she came to help us run the fi rst practical course. I remem- ber it like it was yesterday: We were passaging mouse embryonic fi broblasts (MEFs) together the fi rst day we met, and after centrifuging the cells, we looked at the pellet and at the same time said “fi ve million.” I knew straight away we were talking the same language. We have been collaborating since then, working with stem cells, very often looking at the same problem from a different angle. We complement each other’s knowledge. After looking in the micro- scope and watching stem cells “being” for more than thirteen years, we now feel we have a story to tell. It is what this book is about: the story of stem cells in culture. Step by step, day after day, month after month, thirteen years on a story that is really fascinating, intriguing, and sometimes frustrating. We were always passionate and very often emotional about it. I am absolutely sure anyone going into stem cell research should feel the same: passionate and determined. I n 2006, I joined the Department of Biochemical Engineering at University College London. Every day in the lab, I mentor undergraduate, master, and PhD/EngD students, helping them to understand the practical complexity of stem cell research, and I act as an adviser on numerous collaborative research and development programs. With the rest of my time, I manage the Cell Therapy Research Facilities. I run stem cell practicals and tissue culture courses for under- graduate and postgraduate students who very often have only an engineering, mathematical, or physics background. The lack of a stem cell atlas was really clear from the beginning. From day one, I have had all my images printed in a folder, and before going to the tissue culture practical, I would introduce everyone to “stem cells in pictures” fi rst, discussing all the pros and cons. Only after that would we start the practicals, with a basic knowledge. Every partici- pant would have copies of the pictures. We would compare cells in real culture to cells in the images. It was always very productive this way. S ometimes, I found it easy to teach students without any experience in stem cells or even without any tissue culture experience. Most importantly, I found that I was learning along with them. Most of the participants are students with a biochemical engineering background, and their questions are unexpected for a biologist, coming from a totally different angle. Stem cells are very unpredictable. They are challenging but exciting to work with. My students are always surprised by my excitement and enthusiasm when I am talking about stem cells. I am very often surprised myself, and sometimes I wonder why. I guess it happened after I realized that human pluripotent stem cells are like humans: they resemble each other, and at the same time they differ, each in their own way, with their own special character. Lively or moody, easygo- ix