M E T H O D S I N M O L E C U L A R M E D I C I N ETM PPllaacceennttaa aanndd TTrroopphhoobbllaasstt MMeetthhooddss aanndd PPrroottooccoollss VVoolluummee 22 EEddiitteedd bbyy MMiicchhaaeell JJ.. SSooaarreess JJooaann SS.. HHuunntt Placenta and Trophoblast M E T H O D S I N M O L E C U L A R M E D I C I N E™ John M. Walker, SERIES EDITOR 126. Congenital Heart Disease: Molecular Diagnostics, and Protocols, edited by Margaret H. Baron, edited by Mary Kearns-Jonker, 2006 2005 125.Myeloid Leukemia: Methods and Protocols, ed- 104. Stroke Genomics: Methods and Reviews, ited by Harry Iland, Mark Hertzberg, edited by Simon J. Read and David Virley, and Paula Marlton, 2006 2004 124.Magnetic Resonance Imaging: Methods and 103.Pancreatic Cancer: Methods and Protocols, Biologic Applications, edited by Pottumarthi V. edited by Gloria H. Su, 2004 Prasad, 2006 102.Autoimmunity: Methods and Protocols, 123.Marijuana and Cannabinoid Research: edited by Andras Perl, 2004 Methods and Protocols, edited by Emmanuel S. 101.Cartilage and Osteoarthritis: Volume 2, Onaivi, 2006 Structure and In Vivo Analysis, edited by 122.Placenta and Trophoblast: Methods and Protocols: Frédéric De Ceuninck, Massimo Sabatini, and Volume 2, edited by Michael J. Soares and Joan S. Philippe Pastoureau, 2004 Hunt, 2006 100.Cartilage and Osteoarthritis: Volume 1, 121. Placenta and Trophoblast: Methods and Protocols: Cellular and Molecular Tools, edited by Volume 1, edited by Michael J. Soares and Joan S. Massimo Sabatini, Philippe Pastoureau, and Hunt, 2006 Frédéric De Ceuninck, 2004 120.Breast Cancer Research Protocols, edited by 99.Pain Research: Methods and Protocols, Susan A. Brooks and Adrian Harris, 2006 edited by David Z. 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Steward, 2003 Matthias W. Hoffman, 2005 90.Suicide Gene Therapy: Methods and Reviews, 108.Hypertension: Methods and Protocols, edited by Caroline J. Springer, 2004 edited by Jérôme P. Fennell and Andrew 89.The Blood–Brain Barrier: Biology and H. Baker, 2005 Research Protocols, edited by Sukriti Nag, 2003 107.Human Cell Culture Protocols, Second 88.Cancer Cell Culture: Methods and Protocols, Edition, edited by Joanna Picot, 2005 edited by Simon P. Langdon, 2003 106.Antisense Therapeutics, Second Edition, 87.Vaccine Protocols, Second Edition, edited by edited by M. Ian Phillips, 2005 Andrew Robinson, Michael J. Hudson, and 105.Developmental Hematopoiesis: Methods Martin P. Cranage, 2003 M E T H O D S I N M O L E C U L A R M E D I C I N E™ Placenta and Trophoblast Methods and Protocols Volume 2 Edited by Michael J. Soares Institute of Maternal–Fetal Biology Division of Cancer and Developmental Biology Department of Pathology and Laboratory Medicine University of Kansas Medical Center, Kansas City, KS Joan S. Hunt University Distinguished Professor, Vice Chancellor for Research Department of Anatomy and Cell Biology University of Kansas Medical Center, Kansas City, KS © 2006 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512 www.humanapress.com All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Molecular BiologyTM is a trademark of The Humana Press Inc. All papers, comments, opinions, conclusions, or recommendations are those of the author(s), and do not necessarily reflect the views of the publisher. This publication is printed on acid-free paper. ∞ ANSI Z39.48-1984 (American Standards Institute) Permanence of Paper for Printed Library Materials. Cover illustration: Background: Figure 5 from Chapter 11 (Volume 1), “Mouse Trophoblast Stem Cells” by J. Quinn et al. Foreground: Figure 2 from Chapter 9 (Volume 2), “Macrophage–Trophoblast Interactions” by G. Mor et al. Cover design by Patricia F. Cleary. 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The fee code for users of the Transactional Reporting Service is: [1-58829-608-3/05 $30.00 ]. eISBN 1-59259-989-3 Printed in the United States of America. 10 9 8 7 6 5 4 3 2 1 Library of Congress Cataloging in Publication Data Placenta and trophoblast: methods and protocols / edited by Michael J. Soares and Joan S. Hunt. p. ; cm. — (Methods in molecular medicine ; 121-122) Includes bibliographical references and index. ISBN 1-58829-404-8 (alk. paper) — ISBN 1-58829-608-3 (alk. paper) 1. Placenta. 2. Molecular biology. [DNLM: 1. Placenta. 2. Molecular Biology. WQ 212 P6974 2005] I. Soares, Michael J. II. Hunt, Joan S. III. Series. QP281.P5435 2005 612.6’3—dc22 2005006428 Preface The aim of the two-volume set of Placenta and Trophoblast: Methods and Protocols is to offer contemporary approaches for studying the biology of the placenta. The chapters contained herein also address critical features of the female organ within which the embryo is housed, the uterus, and some aspects of the embryo–fetus itself, particularly those of common experimental animal models. In keeping with the organization used effectively in other volumes in this series, each chapter has a brief introduction followed by a list of required items, protocols, and notes designed to help the reader perform the experi- ments without difficulty. In both volumes, sources of supplies are given and illustrations highlight particular techniques as well as expected outcomes. A key aspect of these volumes is that the contributors are at the forefronts of their disciplines, thus ensuring the accuracy and usefulness of the chapters. Placenta research has progressed rapidly over the past several decades by taking advantage of the technical advances made in other fields. For example, the reader will note that many techniques, such as reverse transcriptase poly- merase chain reaction, northern and western blotting, microarray analyses and in situ hybridization experiments, are routinely used for dissecting a wide range of experimental questions. Protein analysis and functional experiments on tissues and cells that comprise the maternal–fetal interface benefit from studies in endo- crinology, immunology, and developmental biology. These volumes also present new ideas on investigating gene imprinting and gene transfer via viral vectors. In developing these volumes we encountered the problem of how to organize the contents so as to be reader-friendly. Our decision was to subdi- vide in large part by the chronology of pregnancy so that in vivo aspects of implantation come first, followed by in vitro systems of investigation, then protocols for phenotypic analyses of placentas of several species. Special tech- niques mentioned above conclude Volume I. Volume II continues with proto- cols for studying trophoblast invasion, followed by dissection of how invading trophoblast cells might be received by uterine immune cells. Returning to the pla- centa itself, methods for researching trophoblast endocrine and transport functions are followed by a final series of chapters on how placentas adapt to disease. In this latter group, two chapters offer help to investigators interested in animal models of human placental disorders and two address working with the oxygen switches that program gene expression in early pregnancy, a concept entirely unexplored v vi Preface less than a decade ago. The reader is referred to the Introductions in each of the two volumes for a more detailed description of the contents. This project would not have been possible without the contributions of many individuals. We wish to express our gratitude to the contributing authors for their time, effort, creativity, and their willingness to share their knowledge and expertise. Our deep appreciation and gratefulness also goes to Stacy Mc- Clure for her dedicated efforts in maintaining the organization of the manu- scripts and the correspondence between the editors and the authors. During this process the publisher has provided us with helpful guidance and instruc- tion essential for the completion of this effort. Finally, we hope that these volumes are useful and provide a valuable resource for both trainees and established scientists striving to advance our understanding of this unique, entirely essential organ of reproduction. Michael J. Soares Joan S. Hunt Contents Preface ..............................................................................................................v Contributors .....................................................................................................xi Companion Table of Contents for Volume I .................................................xvii Companion CD-ROM ....................................................................................xxi PART I INTRODUCTION 1 Placenta and Trophoblast: Methods and Protocols: Overview II Michael J. Soares and Joan S. Hunt......................................................3 PART II ANALYSIS OF TROPHOBLAST INVASION 2 In Vivo Analysis of Trophoblast Cell Invasion in the Human Robert Pijnenborg, Elizabeth Ball, Judith N. Bulmer, Myriam Hanssens, Stephen C. Robson, and Lisbeth Vercruysse...................................................................11 3 In Vitro Analysis of Trophoblast Invasion John D. Aplin......................................................................................45 4 An In Vitro Model of Trophoblast Invasion of Spiral Arteries Judith E. Cartwright and Mark Wareing.............................................59 PART III ANALYSIS OF UTEROPLACENTAL IMMUNE CELLS AND THEIR FUNCTIONS 5 In Vivo Models for Studying Homing and Function of Murine Uterine Natural Killer Cells B. Anne Croy and Xuemei Xie............................................................77 6 Immune and Trophoblast Cells at the Rhesus Monkey Maternal–Fetal Interface Thaddeus G. Golos, Gennadiy I. Bondarenko, Edith E. Breburda, Svetlana V. Dambaeva, Maureen Durning, and Igor I. Slukvin.....93 7 Methods for Isolation of Cells from the Human Fetal–Maternal Interface Anita Trundley, Lucy Gardner, Jacquie Northfield, Chiwen Chang, and Ashley Moffett..............................................109 vii viii Contents 8 In Vitro Models for Studying Human Uterine and Placental Macrophages Ramsey H. McIntire, Margaret G. Petroff, Teresa A. Phillips, and Joan S. Hunt...........................................................................123 9 Macrophage–Trophoblast Interactions Gil Mor, Shawn L. Straszewski-Chavez, and Vikki M. Abrahams....149 10 Methods for Evaluating Histocompatibility Antigen Gene Expression in the Baboon Daudi K. Langat, Asgerally T. Fazleabas, and Joan S. Hunt.............165 11 Analysis of the Soluble Isoforms of HLA-G: mRNAs and Proteins Judith L. Pace, Pedro J. Morales, Teresa A. Phillips, and Joan S. Hunt...........................................................................181 PART IV ANALYSIS OF PLACENTAL FUNCTION: TRANSPORT AND ENDOCRINOLOGY 12 In Vivo Techniques for Studying Fetoplacental Nutrient Uptake, Metabolism, and Transport Timothy R. H. Regnault and William W. Hay, Jr..............................207 13 In Vitro Models for Studying Trophoblast Transcellular Transport Claudia J. Bode, Hong Jin, Erik Rytting, Peter S. Silverstein, Amber M. Young, and Kenneth L. Audus.....................................225 14 In Vitro Methods for Studying Human Placental Amino Acid Transport: Placental Plasma Membrane Vesicles Jocelyn D. Glazier and Colin P. Sibley.............................................241 15 In Vitro Methods for Studying Human Placental Amino Acid Transport: Placental Villous Fragments Susan L. Greenwood and Colin P. Sibley.........................................253 16 Methods for Investigating Placental Fatty Acid Transport Yan Xu, Thomas J. Cook, and Gregory T. Knipp..............................265 17 Heterologous Expression Systems for Studying Placental Transporters Vadivel Ganapathy, You-Jun Fei, and Puttur D. Prasad...................285 18 Analysis of Trophoblast Giant Cell Steroidogenesis in Primary Cultures Noa Sher and Joseph Orly................................................................301 19 Establishment of an ELISA for the Detection of Native Bovine Pregnancy-Associated Glycoproteins Secreted by Trophoblast Binucleate Cells Jonathan A. Green and R. Michael Roberts......................................321 Contents ix 20 Alkaline Phosphatase Fusion Proteins as Tags for Identifying Targets for Placental Ligands Heiner Müller and Michael J. Soares................................................331 21 Bacterial Expression of Prolactin Family Proteins Arieh Gertler.....................................................................................341 22 Analysis of Placental Regulation of Hematopoiesis Beiyan Zhou and Daniel I. H. Linzer................................................355 23 Methods for Studying Interferon Tau Stimulated Genes Fuller W. Bazer and Thomas E. Spencer...........................................367 PART V ANALYSIS OF PLACENTA ADAPTATION TO DISEASE 24 Reduced Uterine Perfusion Pressure (RUPP) Model for Studying Cardiovascular–Renal Dysfunction in Response to Placental Ischemia Joey P. Granger, B. Babbette D. LaMarca, Kathy Cockrell, Mona Sedeek, Charles Balzi, Derrick Chandler, and William Bennett.....................................................................383 25 In Vivo Rat Model of Preeclampsia S. Ananth Karumanchi and Isaac E. Stillman....................................393 26 A Novel Mouse Model for Preeclampsia by Transferring Activated Th1 Cells into Normal Pregnant Mice Ana Claudia Zenclussen....................................................................401 27 Working with Oxygen and Oxidative Stress In Vitro Graham J. Burton, D. Stephen Charnock-Jones, and Eric Jauniaux..........................................................................413 28 Hypobaric Hypoxia as a Tool to Study Pregnancy-Dependent Responses at the Maternal–Fetal Interface Jennifer K. Ho-Chen, Rupasri Ain, Adam R. Alt, John G. Wood, Norberto C. Gonzalez, and Michael J. Soares.............................427 29 Infection with Listeria monocytogenes as a Probe for Placental Immunological Function Ellen M. Barber, Indira Guleria, and Jeffrey W. Pollard..................435 Index ............................................................................................................443