Cell Fusion: Overviews and Methods

261 Pages·2015·8.609 MB·English

by  Kurt Pfannkuche

Methods in
Molecular Biology 1313
Kurt Pfannkuche Editor
Cell Fusion
Overviews and Methods
Second Edition
M M B
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life and Medical Sciences
University of Hertfordshire
Hat fi eld, Hertfordshire, AL10 9AB, UK
For further volumes:
http://www.springer.com/series/7651

Cell Fusion

Overviews and Methods
Second Edition
Edited by
Kurt Pfannkuche
Center for Physiology and Pathophysiology, University of Cologne, Cologne, Germany
Editor
Kurt P fannkuche
Center for Physiology and Pathophysiology
University of Cologne
Cologne, Germany
ISSN 1064-3745 ISSN 1940-6029 (electronic)
Methods in Molecular Biology
ISBN 978-1-4939-2702-9 ISBN 978-1-4939-2703-6 (eBook)
DOI 10.1007/978-1-4939-2703-6
Library of Congress Control Number: 2015938703
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Prefa ce
The event of cell-cell fusion is involved in a variety of physiological, but importantly also of
pathophysiological processes. As a prominent example, the formation of skeletal muscle
fi bers depends on the fusion of myoblast cells to form giant syncytia. This book addresses
this fascinating process that takes place during muscle development and regeneration with
several chapters highlighting methods to analyze cell fusion in skeletal muscle.
Critically, cell fusion represents an indispensable mechanism in various processes of
development and regeneration. Furthermore, the process of cell fusion may rejuvenate
aging or damaged cells upon their fusion with certain stem or progenitor cells. These
aspects are covered by a selection of protocols that address the fusion process in a variety of
cell types beyond skeletal muscle tissue. High defi nition methods are presented to detect
fusion events in vitro and in vivo and supposed to help scientists in order to gain deeper
insights into the underlying mechanisms of these cell biologically highly important and
relevant processes.
This volume intends to support the reader specifi cally with a collection of state-of-the-
art protocols that allow for the detection and analysis of fusion events in various cell types
in vitro and in vivo. In addition, the provided selection of protocols aims to facilitate the
future highly relevant process of engineering cells for specifi c purposes, such as generation
of transgenic embryos and development of hybrid cells, in order to tackle specifi c tasks in
cell biology and medicine. The book closes with a chapter on mitochondrial fusion as a
prominent process of subcellular fusion processes with a dominant importance for mito-
chondrial function in health and disease—highlighting that not solely cell fuse, but also cell
organelles possess this central biological function in order to fulfi ll their properties within a
healthy organism.
This book would not have been possible without the contribution of many outstanding
experts in the fi eld of cell fusion that agreed to share their knowledge and laboratory pro-
tocols with the readers of this edition. Therefore, I would like to express my honest thank-
fulness to all contributing authors who spend a lot of time and effort to record their
protocols and support me with the project. I am also thankful to John Walker for giving me
the chance to edit this volume of Methods in Molecular Biology.
Cologne, Germany K urt P fannkuche
v

Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . i x
1 State of the Art in Cell–Cell Fusion. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Lena W illkomm and Wilhelm Bloch
2 F usion in Cancer: An Explanatory Model for Aneuploidy, Metastasis
Formation, and Drug Resistance. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Marieke M ohr , Kurt S . Zaenker , and Thomas Dittmar
3 M ouse Embryos’ Fusion for the Tetraploid Complementation Assay. . . . . . . . 4 1
Marina Gertsenstein
4 G eneration of Mouse Chimeras with High Contribution of Tetraploid
Embryonic Stem Cells and Embryonic Stem Cell-Fibroblast Hybrid Cells. . . . 61
Natalia M. M atveeva , Elena A. K izilova , and Oleg L. Serov
5 Microfluidic Systems for Cell Pairing and Fusion. . . . . . . . . . . . . . . . . . . . . . . 7 3
Burak D ura and Joel Voldman
6 C hromosome Tracking in Fused Cells by Single Nucleotide
Polymorphisms. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9 5
Azra Fatima , Jürgen H escheler , and Tomo Šarić
7 F usion of Mesenchymal Stem Cells and Islet Cells for Cell Therapy. . . . . . . . . 1 07
Shoichiro S umi and G oichi Y anai
8 D etection of Fusion Events in Mammalian Skeletal Muscle . . . . . . . . . . . . . . . 1 15
Frank S uhr
9 F ISH Detection of X and Y Chromosomes in Combination
with Immunofluorescence to Study Contribution of Transplanted Cells
to Skeletal Muscle Fibers. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
Anna Strömberg and Monika Jansson
10 U sing Phosphatidylserine Exposure on Apoptotic Cells
to Stimulate Myoblast Fusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 41
Amelia E . H ochreiter-Hufford , S anja A randjelovic ,
and Kodi S . Ravichandran
11 T racing Myoblast Fusion in Drosophila Embryos by Fluorescent
Actin Probes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 149
Shruti Haralalka and S usan M . A bmayr
12 A nalyzing Cell Fusion Events Within the Central Nervous System
Using Bone Marrow Chimerism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1 65
Kevin K emp and K elly Hares
13 Cell Fusion Between Dendritic Cells and Whole Tumor Cells . . . . . . . . . . . . . 1 85
Shigeo K oido and Jianlin G ong
vii
viii Contents
14 Membrane Nanotube Formation in Osteoclastogenesis. . . . . . . . . . . . . . . . . . 1 93
Toshio K ukita , A kira Takahashi , J ing-Qi Zhang , and Akiko Kukita
15 Modified Adherence Method (MAM) for Electrofusion
of Anchorage-Dependent Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 203
Marko U šaj and M aša Kandušer
16 F RET in the Analysis of In Vitro Cell–Cell Fusion by Flow Cytometry . . . . . . 2 17
Guillermo Gómez-Icazbalceta , M irna B erenice R uiz-Rivera ,
Edmundo L amoyi , and Leonor H uerta
17 D ual Split Protein (DSP) Assay to Monitor Cell–Cell Membrane Fusion. . . . . 2 29
Shuhei N akane and Z ene M atsuda
18 Photoconvertible Fluorescent Protein-Based Live Imaging
of Mitochondrial Fusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 37
Geoffrey C. O wens and David B . E delman
I ndex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 47
Contributors
SUSAN M . A BMAYR • Stowers Institute for Medical Research , K ansas City, M O, USA ;
Department of Anatomy and Cell Biology , U niversity of Kansas School of Medicine ,
Kansas City, K S , U SA
SANJA ARANDJELOVIC • Department of Microbiology, Immunology, and Cancer Biology ,
University of Virginia , Charlottesville, VA , U SA
WILHELM B LOCH • Department of Molecular and Cellular Sport Medicine, Institute
of Cardiovascular Research and Sport Medicine , G erman Sport University Cologne ,
Cologne , G ermany
THOMAS D ITTMAR • Institute of Immunology and Experimental Oncology, Centre
for Biomedical Education and Research, U niversity of Witten/Herdecke , W itten , Germany
BURAK DURA • Research Laboratory of Electronics, M assachusetts Institute of Technology
(MIT) , Cambridge, M A , U SA ; E lectrical Engineering and Computer Science
Department , M IT , C ambridge, MA, USA
DAVID B. EDELMAN • Department of Psychological Sciences, U niversity of San Diego ,
San Diego, CA , U SA
AZRA F ATIMA • Center for Physiology and Pathophysiology, Institute for Neurophysiology,
Medical Faculty, U niversity of Cologne , C ologne, Germany
MARINA GERTSENSTEIN • Toronto Centre for Phenogenomics (TCP), Lunenfeld-Tanenbaum
Research Institute, Mount Sinai Hospital , Toronto , O N , C anada
GUILLERMO GÓMEZ-ICAZBALCETA • Department of Immunology, Instituto de Investigaciones
Biomédicas, U niversidad Nacional Autónoma de México , D istrito Federal, M éxico
JIANLIN GONG • Department of Medicine, Boston University School of Medicine , Boston,
MA , U SA
SHRUTI HARALALKA • Stowers Institute for Medical Research , K ansas City, M O, USA
KELLY H ARES • Multiple Sclerosis and Stem Cell Group, School of Clinical Sciences,
University of Bristol , B ristol, UK
JÜRGEN HESCHELER • Center for Physiology and Pathophysiology, Institute for
Neurophysiology, Medical Faculty, University of Cologne, Cologne, Germany
AMELIA E . HOCHREITER-HUFFORD • Department of Microbiology, Immunology, and Cancer
Biology , U niversity of Virginia , C harlottesville, V A , U SA
LEONOR HUERTA • Department of Immunology, Instituto de Investigaciones Biomédicas,
Universidad Nacional Autónoma de México , Distrito Federal, M éxico
MONIKA JANSSON • Department of Medicine, Center for Hematology and Regenerative
Medicine, Karolinska Institutet, K arolinska University Hospital Huddinge , S tockholm ,
Sweden
MAŠA K ANDUŠER • Faculty of Electrical Engineering, University of Ljubljana ,
Ljubljana, S lovenia
KEVIN KEMP • Multiple Sclerosis and Stem Cell Group, School of Clinical Sciences,
University of Bristol , B ristol, UK
ix