Protocols for Neural Cell Culture: Fourth Edition

395 Pages·2010·5.06 MB·English

by  Sharon A. Louis, Brent A. Reynolds (auth.), Laurie C. Doering (eds.)

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Protocols for Neural Cell Culture
Edited by
Laurie C. Doering, PhD
McMaster University, Hamilton, ON, Canada
Fourth Edition
Editor
Laurie C. Doering
Department of Pathology &
Molecular Medicine
McMaster University
1200 Main Street West
Health Sciences Centre
Hamilton ON L8N 3Z5
Canada
Preface
Ross G. Harrison’s founding experiments with nerve cell cultures between 1907 and 1910
were motivated by the same concerns of Santiago Ramo´n y Cajal centered on the “Neu-
ron Doctrine.” Cajal used impressive silver impregnations in chick spinal cord to illustrate
axonal growth in vivo. Harrison, using pieces of frog neural tube nourished in lymph
within a depression slide, solved the problems of his predecessors and devised a repro-
ducible method of tissue culture experimentation. For the first time, the outgrowth rates
of individual fibers and their growth cones were observed in real time under the micro-
scope. In essence, Harrison established a direct in vitro test to prove Cajal’s axon out-
growth theory. Harrison overcame basic tissue culture problems and created a culture
technique others could follow. Since the beginning of nervous system tissue culture with
Ross Harrison’s vision, now just over 100 years ago, the scientific community has estab-
lished numerous protocols to generate the current wide variety of cell and tissue culture
technology.
The rapid growth of neuroscience during the 1980s and a highly acclaimed, intensive
tissue culture course held in Saskatoon, Saskatchewan, for several years formed the concept
for this protocol series in neural tissue culture. In 1992, Dr. Sergey Fedoroff (University of
Saskatchewan) founded this series with the publication of the first edition. The textbook
continued to grow in popularity with subsequent editions, and the book is now established
as a popular neuroscience protocol reference in numerous laboratories throughout the
world.
The fourth edition of Protocols for Neural Cell Culture represents a turning point
in this series from editorial and content perspectives. Since the publication of the third
edition in 2001, neuroscience has continued to expand with discoveries that utilize tissue
culture methodologies. The refinement of existing protocols and the emergence of new
techniques and culture media formulations are linked with advances in neuroscience. This
edition includes three chapters from leading companies who specialize in neural tissue
culture biotechnology and contribute significantly to the products used by many scientists.
Updates on the experimental procedures for many of the classical tissue culture
preparations in neuroscience are highlighted. In view of the implications for regenera-
tive medicine, methods to grow and expand embryonic and adult neural stem cells are
included in this edition. This volume covers the isolation, expansion, and cryopreserva-
tion of neural tissue from mouse, rat, and human sources. Immunocytochemistry is a
subcomponent of many chapters as it continues to be a solid method to identify cells at
developmental points within specific lineages. The basic techniques for establishing speci-
fied neuronal and glial preps are complimented by many new chapters including methods
to assess aspects of cell function (calcium imaging) and cell death.
I am privileged to have the support from Dr. Fedoroff (a great neuroscience mentor)
to continue the Editorship of this collection. I am very grateful to all the authors for their
commitment and time. Their dedication has produced a text with exceptional quality and
v
vi Preface
coverage in tissue culture protocols for today’s neuroscience. A special thanks to Sharon
Ralph for countless hours of editorial assistance, management, and organization in the
production of this book.
Hamilton, ON Laurie C. Doering
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v
Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
1. Neurosphere and Neural Colony-Forming Cell Assays . . . . . . . . . . . . . . 1
Sharon A. Louis and Brent A. Reynolds
2. Directed Neuronal Differentiation of Embryonic and Adult-Derived
Neurosphere Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Marcos R. Costa, Ravi Jagasia, and Benedikt Berninger
3. Culture and Differentiation of Human Neural Stem Cells . . . . . . . . . . . . 51
Soojung Shin and Mohan Vemuri
4. Neural Differentiation of Human Embryonic Stem Cells . . . . . . . . . . . . . 75
Mirella Dottori, Alice Pe´bay, and Martin F. Pera
5. Isolation and Culture of Primary Human CNS Neural Cells . . . . . . . . . . . 87
Manon Blain, Veronique E. Miron, Caroline Lambert,
Peter J. Darlington, Qiao-Ling Cui, Philippe Saikali, V. Wee Yong,
and Jack P. Antel
6. Bioengineering Protocols for Neural Precursor Cell Expansion . . . . . . . . . . 105
Behnam A. Baghbaderani, Arindom Sen, Michael S. Kallos,
and Leo A. Behie
7. Intracellular Calcium Assays in Dissociated Primary Cortical Neurons . . . . . . 125
Navjot Kaur, David V. Thompson, David Judd,
David R. Piper, and Richard G. Del Mastro
8. Dissociated Hippocampal Cultures . . . . . . . . . . . . . . . . . . . . . . . . 137
Francine Nault and Paul De Koninck
9. Primary Sensory and Motor Neuron Cultures . . . . . . . . . . . . . . . . . . 161
Andrea M. Vincent and Eva L. Feldman
10. Retinal Cell and Tissue Culture . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Francisco L.A.F. Gomes and Michel Cayouette
11. Preparation of Normal and Reactive Astrocyte Cultures . . . . . . . . . . . . . 193
Jean de Vellis, Cristina A. Ghiani, Ina B. Wanner,
and Ruth Cole
12. Oligodendrocyte Progenitor Cell Culture . . . . . . . . . . . . . . . . . . . . . 217
Akiko Nishiyama, Ryusuke Suzuki, Hao Zuo, and Xiaoqin Zhu
13. Isolation of Microglia Subpopulations . . . . . . . . . . . . . . . . . . . . . . 231
Makoto Sawada and Hiromi Suzuki
vii
viii Contents
14. Microglia from Progenitor Cells in Mouse Neopallium . . . . . . . . . . . . . . 241
Sergey Fedoroff and Arleen Richardson
15. Primary Schwann Cell Cultures . . . . . . . . . . . . . . . . . . . . . . . . . . 253
Haesun A. Kim and Patrice Maurel
16. Primary Dissociated Astrocyte and Neuron Co-culture . . . . . . . . . . . . . . 269
Shelley Jacobs and Laurie C. Doering
17. Cerebellar Slice Cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 285
Josef P. Kapfhammer
18. Hippocampal Slice Cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . 299
Jesse E. Hanson, Adrienne L. Orr, Silvia Fernandez-Illescas,
Ricardo A. Valenzuela, and Daniel V. Madison
19. Molecular Substrates for Growing Neurons in Culture . . . . . . . . . . . . . . 313
Saulius Satkauskas, Arnaud Muller, Morgane Roth,
and Dominique Bagnard
20. Guidance and Outgrowth Assays for Embryonic Thalamic Axons . . . . . . . . 329
Alexandre Bonnin
21. Detection of Cell Death in Neuronal Cultures . . . . . . . . . . . . . . . . . . 343
Sean P. Cregan
22. Live Imaging of Neural Cell Functions . . . . . . . . . . . . . . . . . . . . . . 353
Sabine Bavamian, Eliana Scemes, and Paolo Meda
23. Tissue Culture Procedures and Tips . . . . . . . . . . . . . . . . . . . . . . . . 375
Arleen Richardson and Sergey Fedoroff
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 391
Contributors
JACK P. ANTEL • Neuroimmunology Unit, Montreal Neurological Institute, Montreal, QC,
Canada
BEHNAM A. BAGHBADERANI • Pharmaceutical Production Research Facility (PPRF),
Schulich School of Engineering, University of Calgary, Calgary, AB, Canada
DOMINIQUE BAGNARD • Physiopathologie du Syste`me Nerveux, INSERM U575, Stras-
bourg, France
SABINE BAVAMIAN • Department of Cell Physiology and Metabolism, University of Geneva
Medical School, Gene`ve, Switzerland
LEO A. BEHIE • Pharmaceutical Production Research Facility (PPRF), Schulich School
of Engineering, University of Calgary, Calgary, AB, Canada
BENEDIKT BERNINGER • Department of Physiological Genomics, Institute of Physiology,
Ludwig-Maximilians University Munich, Schillerstrasse 46, D-80336 Munich, Germany
MANON BLAIN • Neuroimmunology Unit, Montreal Neurological Institute, Montreal, QC,
Canada
ALEXANDRE BONNIN • Zilkha Neurogenetic Institute, Keck School of Medicine of
University of Southern California, 1501 San Pablo Street, Room 105, Los Angeles, CA
90089-2821
MICHEL CAYOUETTE • Cellular Neurobiology Laboratory, Institut de Recherches Clin-
iques de Montre´al, Montre´al, QC, Canada; De´partement de Me´decine, Universite´ de
Montre´al, Montre´al, QC, Canada; Division of Experimental Medicine and Anatomy and
Cell Biology Department, McGill University, Montre´al, QC, Canada
RUTH COLE • Mental Retardation Research Center, Semel Institute for Neuroscience and
Human Behavior, David Geffen School of Medicine at UCLA, Neuroscience Research
Building, Los Angeles, CA, USA
MARCOS R. COSTA • Edmond and Lily Safra International Institute of Neuroscience of
Natal, Natal, RN 59066-060; Universidade Federal do Rio Grande do Norte, Brazil
SEAN P. CREGAN • Robarts Research Institute, London, ON, Canada
QIAO-LING CUI • Neuroimmunology Unit, Montreal Neurological Institute, Montreal,
QC, Canada
PETER J. DARLINGTON • Neuroimmunology Unit, Montreal Neurological Institute,
Montreal, QC, Canada
LAURIE C. DOERING • Department of Pathology and Molecular Medicine, McMaster
University, Hamilton, ON, Canada
MIRELLA DOTTORI • Centre for Neuroscience and Department of Pharmacology, The
University of Melbourne, Parkville, VIC, Australia
SERGEY FEDOROFF • University of Saskatchewan, Saskatoon, SK, Canada
EVA L. FELDMAN • Department of Neurology, University of Michigan, Ann Arbor, MI,
USA
SILVIA FERNANDEZ-ILLESCAS • Stanford University, Stanford, CA, USA
ix
x Contributors
CRISTINA A. GHIANI • Mental Retardation Research Center, Semel Institute for Neuro-
science and Human Behavior, David Geffen School of Medicine at UCLA, Neuroscience
Research Building, Los Angeles, CA, USA
FRANCISCO L.A.F. GOMES • Cellular Neurobiology Laboratory, Institut de Recherches
Cliniques de Montre´al, Montre´al, QC, Canada
JESSE E. HANSON • Stanford University, Stanford, CA, USA
SHELLEY JACOBS • Department of Pathology and Molecular Medicine, McMaster
University, Hamilton, ON, Canada
RAVI JAGASIA • Junior Research Group Adult Neurogenesis and Neural Stem Cells, Insti-
tute of Developmental Genetics, Helmholtz Zentrum Mu¨nchen, German Research Center
for Environmental Health, Munich-Neuherberg, Germany
DAVID JUDD • Invitrogen Corporation, Grand Island, NY, USA
MICHAEL S. KALLOS • Pharmaceutical Production Research Facility (PPRF), Schulich
School of Engineering, University of Calgary, Calgary, AB, Canada
JOSEF P. KAPFHAMMER • Anatomisches Institut, Universita¨t Basel, Basel, Switzerland
NAVJOT KAUR • Invitrogen Corporation, Frederick, MD, USA
HAESUN A. KIM • Department of Biological Sciences, Rutgers University, Newark, NJ,
USA
PAUL DE KONINCK • Cellular Neurobiology Division, Centre de Recherche, Universite´
Laval Robert Giffard, Que´bec, QC, Canada
CAROLINE LAMBERT • Neuroimmunology Unit, Montreal Neurological Institute,
Montreal, QC, Canada
SHARON A. LOUIS • STEMCELL Technologies Inc., Vancouver, BC, Canada
DANIEL V. MADISON • Stanford University, Stanford, CA, USA
RICHARD G. DEL MASTRO • Invitrogen Corporation, Grand Island, NY, USA
PATRICE MAUREL • Department of Cell Biology, Smilow Neuroscience Program, New York
University School of Medicine, New York, NY, USA
PAOLO MEDA • Department of Cell Physiology and Metabolism, University of Geneva
Medical School, Gene`ve, Switzerland
VERONIQUE E. MIRON • Neuroimmunology Unit, Montreal Neurological Institute,
Montreal, QC, Canada
ARNAUD MULLER • Physiopathologie du Syste`me Nerveux, INSERM U575, Strasbourg,
France
FRANCINE NAULT • Cellular Neurobiology Division, Centre de Recherche, Universite´
Laval Robert Giffard, Que´bec, QC, Canada
AKIKO NISHIYAMA • Department of Physiology and Neurobiology, University of
Connecticut, Storrs, CT, USA; University of Connecticut Stem Cell Institute, University
of Connecticut, Storrs, CT, USA
ADRIENNE L. ORR • Stanford University, Stanford, CA, USA
ALICE PE´BAY • Centre for Neuroscience and Department of Pharmacology, The University
of Melbourne, Parkville, VIC, Australia
MARTIN F. PERA • Center for Stem Cell and Regenerative Medicine, University of Southern
California, Los Angeles, CA, USA
DAVID R. PIPER • Invitrogen Corporation, Madison, WI, USA
BRENT A. REYNOLDs • Department of Neurosurgery, McKnight Brain Institute,
University of Florida, Gainesville, FL, USA
ARLEEN RICHARDSON • University of Saskatchewan, Saskatoon, SK, Canada
Contributors xi
MORGANE ROTH • Physiopathologie du Syste`me Nerveux, INSERM U575, Strasbourg,
France
PHILIPPE SAIKALI • Neuroimmunology Unit, Montreal Neurological Institute, Montreal,
QC, Canada
SAULIUS SATKAUSKAS • Physiopathologie du Syste`me Nerveux, INSERM U575, Stras-
bourg, France; Department of Biology, Vytautas Magnus University, Kaunas, Lithuania
MAKOTO SAWADA • Department of Brain Function, Research Institute of Environmental
Medicine, Nagoya University, Nagoya, Japan
ELIANA SCEMES • The Dominick P. Purpura Department of Neuroscience, Albert Einstein
College of Medicine, Bronx, NY, USA
ARINDOM SEN • Pharmaceutical Production Research Facility (PPRF), Schulich School of
Engineering, University of Calgary, Calgary, AB, Canada
SOOJUNG SHIN • Department of Stem Cell and Regenerative Medicine, Invitrogen Inc.,
Frederick, MD, USA
HIROMI SUZUKI • Department of Brain Function, Research Institute of Environmental
Medicine, Nagoya University, Nagoya, Japan
RYUSUKE SUZUKI • Department of Physiology and Neurobiology, University of Connecti-
cut, Storrs, CT, USA
DAVID V. THOMPSON • Invitrogen Corporation, Madison, WI, USA
RICARDO A. VALENZUELA • Stanford University, Stanford, CA, USA
JEAN DE VELLIS • Mental Retardation Research Center, Semel Institute for Neuroscience
and Human Behavior, David Geffen School of Medicine at UCLA, Neuroscience Research
Building, Los Angeles, CA, USA
MOHAN VEMURI • Department of Stem Cell and Regenerative Medicine, Invitrogen Inc.,
Frederick, MD, USA
ANDREA M. VINCENT • Department of Neurology, University of Michigan, Ann Arbor,
MI, USA
INA B. WANNER • Mental Retardation Research Center, Semel Institute for Neuroscience
and Human Behavior, David Geffen School of Medicine at UCLA, Neuroscience Research
Building, Los Angeles, CA, USA
V. WEE YONG • Departments of Clinical Neurosciences and Oncology, University of Cal-
gary, Calgary, AB, Canada
XIAOQIN ZHU • Department of Physiology and Neurobiology, University of Connecticut,
Storrs, CT, USA
HAO ZUO • Department of Physiology and Neurobiology, University of Connecticut, Storrs,
CT, USA