M e t h o d s i n M o l e c u l a r B i o l o g y ™

Series EditorJohn M. Walker

School of Life SciencesUniversity of Hertfordshire

Hatfield, Hertfordshire, AL10 9AB, UK

For other titles published in this series, go to www.springer.com/series/7651

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Adipose-Derived Stem Cells

Methods and Protocols

Edited by

Jeffrey M. Gimble

Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA, USA

Bruce A. Bunnell

Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA

EditorsJeffrey M. Gimble, MD, Ph.D.Stem Cell Biology LaboratoryPennington Biomedical Research CenterBaton Rouge, LAUSAjeffrey.gimble@pbrc.edu

Bruce A. Bunnell, Ph.D.Center for Stem Cell Research and Regenerative MedicineTulane University School of MedicineNew Orleans, LAUSAbbunnell@tulane.edu

ISSN 1064-3745 e-ISSN 1940-6029ISBN 978-1-61737-959-8 e-ISBN 978-1-61737-960-4DOI 10.1007/978-1-61737-960-4Springer New York Dordrecht Heidelberg London

© Springer Science+Business Media, LLC 2011All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden.The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights.While the advice and information in this book are believed to be true and accurate at the date of going to press, 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

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v

Preface

During the past decade, a wide range of scientific disciplines have “found” adipose-derived stem/stromal cells (ASC) as a tool for research and discovery. This is reflected by the exponential growth in the number of publications and citations noted by ISI Web of Science (Thomson-Reuters) (Fig. 1). However, the cells we now recognize as “ASC” were initially identified as “preadipocytes” in seminal studies initiated by Martin Rodbell and his col-leagues during the mid-1960s. His laboratory was among the first to use collagenase digestion to separate mature adipocytes from the heterogeneous stromal vascular fraction (SVF) cells in rat fat pads. Since then, primary cultures of preadipocytes have been utilized by endocrinologists and physiologists to perform in vitro analyses of adipose tissue metab-olism. By the late 1990s, investigators at the University of Pittsburgh, University of California – Los Angeles, Zen-Bio, Inc., and other institutions had begun to draw parallels between the ASC and the more thoroughly characterized bone marrow-derived mesen-chymal stem/stromal cells (BMSC). Both ASC and BMSC displayed multipotent differ-entiation potential in vitro, with the ability to form bone, cartilage, hematopoietic supporting cells, and muscle, in addition to adipocytes. Later studies would document that they also share similar immunophenotypic and immunomodulatory characteristics. Unlike bone marrow, subcutaneous adipose tissue is relatively accessible to harvest, abun-dant, and located in a site that patients are willing, indeed eager, to have biopsied. Consequently, the demand for ASC has grown; they quickly have become the stem cell of choice for many tissue-engineering and regenerative medical projects. Bioengineers, clini-cians, entrepreneurs, and research scientists recognized the need to coordinate this emerg-ing field and founded the International Federation of Adipose Therapeutics and Science (IFATS) in 2002. IFATS was envisioned as a society that would promote the free exchange of information and knowledge relating to ASC. In its efforts to provide standardization across laboratories, one of the first steps of IFATS was to establish the acronym “ASC” to describe the collagenase-digested and culture-expanded adipose cell populations. Prior to reaching this consensus in 2004, the nomenclature had been cluttered with multiple terms that led to confusion for newcomers in the field. In this volume of Methods in Molecular Biology, we have solicited defined and established protocols from leaders in the field, including many IFATS members. While much of the emphasis is placed on human ASC, additional small and large animal species are included. The chapters are organized around approaches spanning the discovery, preclinical, and clinical processes. While they are designed to be accessible to new students, we hope that they will serve as a reference text for established investigators as well.

The editors have many people to thank for their contributions to this work. First and foremost are the many authors who agreed to take time from their busy schedules and prepare chapters on selected topics (with only a minimum of bribes and arm twisting!). Second is Professor John Walker, Series Editor, whose sage advice and wisdom were greatly appreciated. Third are the members of the Stem Cell Biology Laboratory, Pennington

vi Preface

Biomedical Research Center, and the Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, for their critiques and insights. Fourth, is Ms. Laura Dallam for her outstanding administrative and editorial assistance. And finally, for sacrificing many evenings and weekends, are our family members: Emma, Jesse, Megan, Paula, and Xiying.

Baton Rouge, LA Jeffrey M. GimbleNew Orleans, LA Bruce A. Bunnell

Fig. 1 The ISI Web of Knowledge annual publications and citations for “adipose stem cells”

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vii

Contents

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vContributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

Part I IsolatIon Methods

1 Isolation and Culture of Rhesus Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . 3Christine Gagliardi and Bruce A. Bunnell

2 Isolation of Human Adipose-Derived Stem Cells from Lipoaspirates . . . . . . . . . . . 17Gang Yu, Z. Elizabeth Floyd, Xiying Wu, Yuan-Di C. Halvorsen, and Jeffrey M. Gimble

3 Isolation of Murine Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . 29Gang Yu, Xiying Wu, Gail Kilroy, Yuan-Di C. Halvorsen, Jeffrey M. Gimble, and Z. Elizabeth Floyd

4 In Vitro Adult Rat Adipose Tissue-Derived Stromal Cell Isolation and Differentiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37Mandi J. Lopez and Nakia D. Spencer

5 In Vitro Adult Canine Adipose Tissue-Derived Stromal Cell Growth Characteristics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47Nakia D. Spencer and Mandi J. Lopez

6 Adipogenic Differentiation of Adult Equine Mesenchymal Stromal Cells. . . . . . . . 61Martin A. Vidal and Mandi J. Lopez

7 Isolation and Culture of Porcine Adipose Tissue-Derived Somatic Stem Cells . . . . 77Kellie J. Williams, Robert A. Godke, and Kenneth R. Bondioli

8 Automated Isolation and Processing of Adipose-Derived Stem and Regenerative Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87Kevin C. Hicok and Marc H. Hedrick

Part II PhenotyPIng Methods

9 Methods for the Purification and Characterization of Human Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109Stan Gronthos and Andrew C.W. Zannettino

10 Characterization of Human Adipose-Derived Stem Cells Using Flow Cytometry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121H. Alan Tucker and Bruce A. Bunnell

11 Evaluation of Cellular and Humoral Immune Responses to Allogeneic Adipose-Derived Stem/Stromal Cells . . . . . . . . . . . . . . . . . . . . . . . 133Kevin R. McIntosh

viii Contents

12 Adipose-Derived Stromal Cells: Cytokine Expression and Immune Cell Contaminants . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151Pauline Decaunes, David Estève, Alexia Zakaroff-Girard, Coralie Sengenès, Jean Galitzky, and Anne Bouloumié

13 Gel-Based and Gel-Free Proteomic Technologies . . . . . . . . . . . . . . . . . . . . . . . . . 163Peter Scherp, Ginger Ku, Liana Coleman, and Indu Kheterpal

Part III dIfferentIatIon Methods: In VItro and In VIVo aPPlIcatIons

14 Adipogenic Differentiation of Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . 193Gang Yu, Z. Elizabeth Floyd, Xiying Wu, Teddi Hebert, Yuan-Di C. Halvorsen, Benjamin M. Buehrer, and Jeffrey M. Gimble

15 Three-Dimensional Culture Systems to Induce Chondrogenesis of Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 201Bradley T. Estes and Farshid Guilak

16 Neural Differentiation of Human Adipose Tissue-Derived Stem Cells . . . . . . . . . . 219Ji Min Yu, Bruce A. Bunnell, and Soo-Kyung Kang

17 Osteogenic Differentiation Strategies for Adipose-Derived Mesenchymal Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 233Robert Jan Kroeze, Marlene Knippenberg, and Marco N. Helder

18 Generation of Adipose Stromal Cell-Derived Hepatic Cells . . . . . . . . . . . . . . . . . . 249Joseph C. Ruiz

19 Adipose Stem Cell Differentiation into Smooth Muscle Cells . . . . . . . . . . . . . . . . 261Kacey G. Marra, Candace A. Brayfield, and J. Peter Rubin

20 Endothelial and Cardiac Regeneration from Adipose Tissues. . . . . . . . . . . . . . . . . 269Louis Casteilla, Valérie Planat-Bénard, Stéphanie Dehez, Sandra De Barros, Corinne Barreau, and Mireille André

21 Epithelial Differentiation of Human Adipose-Derived Stem Cells . . . . . . . . . . . . . 289Patrick C. Baer, Martin Brzoska, and Helmut Geiger

22 Screening for Epigenetic Target Genes that Enhance Reprogramming Using Lentiviral-Delivered shRNA . . . . . . . . . . . . . . . . . . . . . . . 299Jong S. Rim, Karen Strickler, Christian Barnes, Lettie Harkins, Jaroslaw Staszkiewicz, Rachel Power, and Kenneth J. Eilertsen

Part IV BIoengIneerIng, ManufacturIng, and clInIcal technologIes

23 Adipogenic Differentiation of Human Adipose-Derived Stem Cells on 3D Silk Scaffolds . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 319Jennifer H. Choi, Evangelia Bellas, Gordana Vunjak-Novakovic, and David L. Kaplan

24 A First Approach for the Production of Human Adipose Tissue-Derived Stromal Cells for Therapeutic Use. . . . . . . . . . . . . . . . . . . . . . . . . 331Philippe Bourin, Julie-Anne Peyrafitte, and Sandrine Fleury-Cappellesso

25 Viral Transduction of Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . 345Patricia A. Zuk

26 Use of Adipose-Derived Stem Cells in High-Throughput Screening to Identify Modulators of Lipogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . 359Y. Reneé Lea-Currie, David J. Duffin, and Benjamin M. Buehrer

ixContents

27 Preservation Protocols for Human Adipose Tissue-Derived Adult Stem Cells . . . . 369Ram Devireddy and Sreedhar Thirumala

28 Soft Tissue Reconstruction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 395J. Peter Rubin and Kacey G. Marra

29 Methods for Analyzing MicroRNA Expression and Function During Osteogenic Differentiation of Human Adipose Tissue-Derived Mesenchymal Stem Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 401Yeon Jeong Kim and Jin Sup Jung

30 The Generation and the Manipulation of Human Multipotent Adipose-Derived Stem Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 419Brigitte Wdziekonski, Tala Mohsen-Kanson, Phi Villageois, and Christian Dani

31 Cell Sheet Technology for Tissue Engineering: The Self-Assembly Approach Using Adipose-Derived Stromal Cells. . . . . . . . . . . . 429Benoît Labbé, Guillaume Marceau-Fortier, and Julie Fradette

32 Differentiation of Adipose-Derived Stem Cells for Tendon Repair . . . . . . . . . . . . . 443A. Cagri Uysal and Hiroshi Mizuno

33 Adipose-Derived Stem Cells for Skin Regeneration . . . . . . . . . . . . . . . . . . . . . . . . 453Hiroshi Mizuno and Masaki Nambu

34 Adipose-Derived Stem Cells for Periodontal Tissue Regeneration . . . . . . . . . . . . . 461Morikuni Tobita and Hiroshi Mizuno

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 471

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xi

Contributors

MIreIlle andré • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie, CNRS, Université de Toulouse, Toulouse, France

PatrIck c. Baer • Division of Nephrology, Department of Internal Medicine III, Goethe-University, Frankfurt, Germany

chrIstIan Barnes • NuPotential, Inc ., Louisiana Emerging Technology Center, Baton Rouge, LA, USA

corInne Barreau • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie, CNRS, Université de Toulouse, Toulouse, France

eVangelIa Bellas • Biomedical Engineering Department, Tissue Engineering Resource Center, Tufts University, Medford, MA, USA

kenneth r. BondIolI • Department of Animal Science, Louisiana State University, Baton Rouge, LA, USA

anne BoulouMIé • Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

PhIlIPPe BourIn • Laboratoire de thérapie cellulaire, EFS-PM, Toulouse, Francecandace a. BrayfIeld • Division of Plastic Surgery, Department of Surgery,

University of Pittsburgh, Pittsburgh, PA, USAMartIn Brzoska • Division of Nephrology, Department of Internal Medicine III,

Goethe-University, Frankfurt, GermanyBenjaMIn M. Buehrer • ZenBio, Inc ., Research Triangle Park, NC, USABruce a. Bunnell • Center for Stem Cell Research and Regenerative Medicine,

Tulane University School of Medicine, New Orleans, LA, USA; Tulane National Primate Research Center, Covington, LA, USA

louIs casteIlla • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie, CNRS, Université de Toulouse, Toulouse, France

jennIfer h. choI • Biomedical Engineering, Tufts University, Medford, MA, USAlIana coleMan • Protein Structural Biology, Pennington Biomedical Research

Center, Louisiana State University System, Baton Rouge, LA, USAchrIstIan danI • Institut de Recherche, Signalisation, Biologie du Développement

et Cancer, Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia-Antipolis, Nice, France

sandra de Barros • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie, CNRS, Université de Toulouse, Toulouse, France

PaulIne decaunes • Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

stéPhanIe dehez • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie, CNRS, Université de Toulouse, Toulouse, France

xii Contributors

raM deVIreddy • Bioengineering Laboratory, Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA, USA

daVId j. duffIn • ZenBio, Inc ., Research Triangle Park, NC, USAkenneth j. eIlertsen • NuPotential, Inc ., Louisiana Emerging Technology Center,

Baton Rouge, LA, USA; Nuclear Reprogramming and Epigenetics Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

daVId estèVe • Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

Bradley t. estes • Division of Orthopaedic Surgery, Department of Surgery, Duke University Medical Center, Durham, NC, USA

sandrIne fleury-caPPellesso • Laboratoire de thérapie cellulaire, EFS-PM, Toulouse, France

z. elIzaBeth floyd • Ubiquitin Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

julIe fradette • LOEX (Laboratoire d’Organogénèse Expérimentale/Experimental Organogenesis Laboratory), Centre de Recherche FRSQ du CHA Universitaire de Québec, Québec QC, Canada; Département de Chirurgie, Université Laval, Québec QC, Canada

chrIstIne gaglIardI • Center for Stem Cell Research and Regenerative Medicine, Department of Pharmacology, Tulane University School of Medicine, New Orleans, LA, USA

jean galItzky • Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

helMut geIger • Division of Nephrology, Department of Internal Medicine III, Goethe-University, Frankfurt, Germany

jeffrey M. gIMBle • Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

roBert a. godke • Department of Animal Science, Louisiana State University, Baton Rouge, LA, USA

stan gronthos • Mesenchymal Stem Cell Group, Department of Haematology, Institute of Medical and Veterinary Science/Hanson Institute and Centre for Stem Cell Research, University of Adelaide, Adelaide, SA, Australia

farshId guIlak • Departments of Surgery and Biomedical Engineering, Duke University Medical Center, Durham, NC, USA

yuan-dI c. halVorsen • Center for Computational and Integrative Biology, Harvard Medical School, Boston, MA, USA

lettIe harkIns • NuPotential, Inc ., Louisiana Emerging Technology Center, Baton Rouge, LA, USA

teddI heBert • Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

Marc h. hedrIck • Cytori Therapeutics, Inc ., San Diego, CA, USAMarco n. helder • Orthopaedic Surgery, VU University Medical Center,

Amsterdam, The Netherlands

xiiiContributors

keVIn c. hIcok • Regenerative Cell Therapeutics Research, Cytori Therapeutics, Inc ., San Diego, CA, USA

jIn suP jung • Department of Physiology, School of Medicine, Pusan National University, Beomeo-ri, Mulgeum-eupYangsan-si Gyeongsangnam-do, Korea

soo-kyung kang • Laboratory of Stem Cell Biology, Department of Biotechnology, School of Veterinary Medicine, Seoul National University, Seoul, Korea

daVId l. kaPlan • Bioengineering Department, Tufts University, Medford, MA, USAIndu kheterPal • Protein Structural Biology, Pennington Biomedical Research

Center, Louisiana State University System, Baton Rouge, LA, USAgaIl kIlroy • Ubiquitin Laboratory, Pennington Biomedical Research Center, Loui-

siana State University System, Baton Rouge, LA, USAyeon jeong kIM • Department of Physiology, School of Medicine, Pusan National

University, Beomeo-ri, Mulgeum-eupYangsan-si Gyeongsangnam-do, KoreaMarlene knIPPenBerg • MOVE/Skeletal Tissue Engineering Group Amsterdam

(STEGA), Amsterdam, The NetherlandsroBert jan kroeze • Department of Orthopaedic Surgery, VU University Medical

Center, Amsterdam, The Netherlands; Department of Oral Cell Biology, Academic Centre for Dentistry Amsterdam (ACTA), Amsterdam, The Netherlands; Universiteit van Amsterdam and Vrije Universiteit, Amsterdam, The Netherlands; MOVE/Skeletal Tissue Engineering Group Amsterdam (STEGA), Amsterdam, The Netherlands

gInger ku • Protein Structural Biology, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

Benoît laBBé • LOEX (Laboratoire d’Organogénèse Expérimentale/Experimental Organogenesis Laboratory), Centre de Recherche FRSQ du CHA Universitaire de Québec, Québec QC, Canada; Département de Chirurgie, Université Laval, Québec QC, Canada

y. reneé lea-currIe • ZenBio, Inc ., Research Triangle Park, NC, USAMandI j. loPez • Laboratory for Equine and Comparative Orthopedic Research,

School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USAguIllauMe Marceau-fortIer • LOEX (Laboratoire d’Organogénèse Expérimentale/

Experimental Organogenesis Laboratory), Centre de Recherche FRSQ du CHA Universitaire de Québec, Québec QC, Canada; Département de Chirurgie, Université Laval, Québec QC, Canada

kacey g. Marra • Division of Plastic Surgery, Department of Surgery, McGowan Institute for Regenerative Medicine, Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA

keVIn r. McIntosh • Cognate BioServices, Inc ., Baltimore, MD, USAhIroshI MIzuno • Department of Plastic and Reconstructive Surgery,

School of Medicine, Juntendo University, Tokyo, Japantala Mohsen-kanson • Institut de Recherche, Signalisation,

Biologie du Développement et Cancer, UMR 6543 Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia-Antipolis, Nice, France

MasakI naMBu • Department of Plastic and Reconstructive Surgery, National Defense Medical College, Tokorozawa, Japan

xiv Contributors

julIe-anne PeyrafItte • Laboratoire de thérapie cellulaire, EFS-PM, Toulouse, FranceValérIe Planat-Bénard • UPS, UMR 5241 Métabolisme, Plasticité et Mitochondrie,

CNRS, Université de Toulouse, Toulouse, Francerachel Power • NuPotential, Inc ., Louisiana Emerging Technology Center,

Baton Rouge LA, USAjong s. rIM • NuPotential, Inc ., Louisiana Emerging Technology Center,

Baton Rouge, LA, USAj. Peter ruBIn • Division of Plastic Surgery, Department of Surgery,

McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA, USA

josePh c. ruIz • Transposagen Biopharmaceuticals, Inc ., Lexington, KY, USAPeter scherP • Proteomics and Metabolomics Core, Pennington Biomedical

Research Center, Louisiana State University System, Baton Rouge, LA, USAcoralIe sengenès • Institut National de la Santé et de la Recherche Médicale

(INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

nakIa d. sPencer • Laboratory for Equine and Comparative Orthopedic Research, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA, USA

jaroslaw staszkIewIcz • NuPotential, Inc ., Louisiana Emerging Technology Center, Baton Rouge, LA, USA

karen strIckler • NuPotential, Inc ., Louisiana Emerging Technology Center, Baton Rouge, LA, USA

sreedhar thIruMala • Bioengineering Laboratory, Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA, USA

MorIkunI toBIta • Office of Surgeon General and Director of Medicine, Maritime Staff Office, Ministry of Defense, Tokyo, Japan

h. alan tucker • Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA

a. cagrI uysal • Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Baskent University, Ankara, Turkey

MartIn a. VIdal • JD Wheat Veterinary Orthopedic Research Laboratory, School of Veterinary Medicine, University of California – Davis, Davis, CA, USA

PhI VIllageoIs • Institut de Recherche, Signalisation, Biologie du Développement et Cancer, UMR 6543 Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia-Antipolis, Nice, France

gordana Vunjak-noVakoVIc • Biomedical Engineering, Vanderbilt Clinic, Columbia University, New York, NY, USA

BrIgItte wdzIekonskI • Institut de Recherche, Signalisation, Biologie du Développement et Cancer, UMR 6543 Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia-Antipolis, Nice, France

kellIe j. wIllIaMs • School of Animal Science, Louisiana State University, Baton Rouge, LA, USA

XIyIng wu • Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

gang yu • Stem Cell Biology Laboratory, Pennington Biomedical Research Center, Louisiana State University System, Baton Rouge, LA, USA

xvContributors

jI MIn yu • Division of Regenerative Medicine, Tulane National Primate Research Center, Covington, LA, USA

aleXIa zakaroff-gIrard • Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Médecine Moléculaire de Rangueil, Université Toulouse III Paul-Sabatier, Toulouse, France

andrew c.w. zannettIno • Myeloma Research Laboratory, Department of Haematology, Institute of Medical and Veterinary Science/Centre for Cancer Biology and the Centre for Stem Cell Research, University of Adelaide, Adelaide SA, Australia

PatrIcIa a. zuk • Regenerative Bioengineering and Repair Laboratory, Division of Plastic Surgery, Department of Surgery, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

Adipose-Derived Stem CellsPrefaceContentsContributors