Human Cell Culture Protocols

432 Pages·2012·6.837 MB·English

by  Christina Philippeos, Robin D. Hughes, Anil Dhawan (auth.), Ragai R. Mitry, Robin D. Hughes (eds.)

M M B ™
ETHODS IN OLECULAR IOLOGY
Series Editor
John M. Walker
School of Life Sciences
University of Hertfordshire
Hatfield, Hertfordshire, AL10 9AB, UK
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Human Cell Culture Protocols
Third Edition
Edited by
Ragai R. Mitry
Institute of Liver Studies, King’s College Hospital, London, UK
Robin D. Hughes
Institute of Liver Studies, King’s College London School of Medicine, London, UK
Editors
Ragai R. Mitry, Ph.D Robin D. Hughes, Ph.D
Institute of Liver Studies Institute of Liver Studies
King’s College Hospital King’s College London
London, UK School of Medicine
[email protected] London, UK
[email protected]
ISSN 1064-3745 e-ISSN 1940-6029
ISBN 978-1-61779-366-0 e-ISBN 978-1-61779-367-7
DOI 10.1007/978-1-61779-367-7
Springer New York Dordrecht Heidelberg London
Library of Congress Control Number: 2011939785
© Springer Science+Business Media, LLC 2012
All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the
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Dedication
My sincere thanks to my mother Elizabeth, sister Antoinette, and brother Nabil for their
love, encouragement, and support throughout my studies and career.
Ragai R. Mitry
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Preface
Welcome to the 3rd edition of H uman Cell Culture Protocols . The 1st edition was published
in 1996 with Gareth Jones as editor and the 2nd edition in 2004 with Joanna Picot as editor.
We hope our edition is a worthy addition to this series. Some might say, “Why do we need
another book on human cell culture?” Human cell culture is not a new topic, but develop-
ment of new molecular techniques and reagents which can be used to investigate cell func-
tion and the intracellular mechanisms responsible make it a continuing requirement. Human
stem cells have become an important area for cell culture but are not described here as they
are dealt with by other volumes. We describe protocols for isolation and culture of a range
of primary cells from human tissues. It is impossible to include all cell types, and, for
instance, we have not included neural cells or cancer cells. Of increasing importance, the
techniques can be adapted to obtain cell cultures from diseased tissues to help understand
the pathogenetic mechanisms involved.
A number of the chapters are new versions of protocols from authors involved in the
previous editions of H uman Cell Culture Protocols , who, with this, are bringing years of
experience of working on particular cell types. There are chapters from authors new to the
series on different cell types. For example, we have included pancreatic cells needed for
basic studies on the pathogenesis of diabetes and also to their application for islet transplan-
tation. There are three chapters on hepatocytes, which is the primary interest of the editors
who work on hepatocyte transplantation. There are also protocols for skin cells, lung cells,
parathyroid cells, gastric cells, renal cells, adipocytes, ovarian cells, bone cells, vascular
smooth muscle cells, vascular endothelial cells, regulatory T cells, and blood mononuclear
cells. There are chapters on new techniques being applied to human cell culture, particularly
the use of biocompatible scaffolds to grow cells, the in vitro use of laser microdissection to
isolate cells from culture, and automated cell culture.
The detailed protocols described in the different chapters make it possible for a worker
with basic cell culture training to prepare cell cultures of the specifi c cell type. It is hoped
that this volume will be of use to scientists in the fi elds of cell biology, gene therapy, and cell
transplantation.
We would like to thank all the contributors for making this book possible.
London Ragai R. Mitry
Robin D. Hughes
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Contents
Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Contributors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
1 Introduction to Cell Culture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Christina Philippeos, Robin D. Hughes, Anil Dhawan,
and Ragai R. Mitry
2 Isolation and Cultivation of Dermal Stem Cells that Differentiate
into Functional Epidermal Melanocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Ling Li, Mizuho Fukunaga-Kalabis, and Meenhard Herlyn
3 Isolation, Cultivation, and Application of Human Alveolar
Epithelial Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Nicole Daum, Anna Kuehn, Stephanie Hein, Ulrich F. Schaefer,
Hanno Huwer, and Claus-Michael Lehr
4 Culture of Parathyroid Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Peyman Björklund and Per Hellman
5 Functional Analysis of Human Islets of Langerhans Maintained
in Culture. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Shanta J. Persaud, Bo Liu, and Peter M. Jones
6 Conversion of Non-endocrine Human Pancreatic Cells
to Insulin-Producing Cells for Treatment of Diabetes. . . . . . . . . . . . . . . . . . . . . . . 73
Min Zhao and Guo Cai Huang
7 Evaluation of Cytochrome P450 Activities in Human
Hepatocytes In Vitro . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
María José Gómez-Lechón, Agustín Lahoz, José V. Castell,
and María Teresa Donato
8 Human Hepatocytes: Isolation, Culture, and Quality Procedures . . . . . . . . . . . . . . 99
Daniel Knobeloch, Sabrina Ehnert, Lilianna Schyschka,
Peter Büchler, Michael Schoenberg, Jörg Kleeff,
Wolfgang E. Thasler, Natascha C. Nussler, Patricio Godoy,
Jan Hengstler, and Andreas K. Nussler
9 Preclinical Testing of Virotherapeutics for Primary and Secondary
Tumors of the Liver . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Martina Zimmermann, Timo Weiland, Michael Bitzer,
and Ulrich M. Lauer
10 Isolation and Functional Studies of Human Fetal Gastric
Epithelium in Primary Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
Pierre Chailler, Jean-François Beaulieu, and Daniel Ménard
11 Isolation, Characterization, and Culture of Normal Human
Intestinal Crypt and Villus Cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Jean-François Beaulieu and Daniel Ménard
ix
x Contents
12 Primary Culture of Human Renal Proximal Tubule Epithelial Cells
and Interstitial Fibroblasts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
Claire C. Sharpe and Mark E.C. Dockrell
13 Glomerular Epithelial and Mesangial Cell Culture
and Characterization. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Heather M. Wilson and Keith N. Stewart
14 Culture of Isolated Human Adipocytes and Isolated Adipose Tissue . . . . . . . . . . . . 203
Kirstin A. Carswell, Mi-Jeong Lee, and Susan K. Fried
15 Primary Culture of Human Adipocyte Precursor Cells:
Expansion and Differentiation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 215
Thomas Skurk and Hans Hauner
16 Primary Culture of Ovarian Cells for Research on Cell Interactions
in the Hormonal Control of Steroidogenesis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 227
Jerzy F. Galas
17 Human Vascular Smooth Muscle Cell Culture . . . . . . . . . . . . . . . . . . . . . . . . . . . . 251
Diane Proudfoot and Catherine Shanahan
18 Culture of Human Endothelial Cells from Umbilical Veins . . . . . . . . . . . . . . . . . . . 265
Richard C.M. Siow
19 Human Peripheral Blood Mononuclear Cell Culture for Flow
Cytometric Analysis of Phosphorylated Mitogen-Activated
Protein Kinases. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275
Athanasios Mavropoulos, Daniel Smyk, Eirini I. Rigopoulou,
and Dimitrios P. Bogdanos
20 Human CD4+CD25highCD127low/neg Regulatory T Cells. . . . . . . . . . . . . . . . . . . . . 287
Haibin Su, Maria Serena Longhi, Pengyun Wang,
Diego Vergani, and Yun Ma
21 Human Chondrocyte Cultures as Models of Cartilage-Specific
Gene Regulation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 301
Miguel Otero, Marta Favero, Cecilia Dragomir, Karim El Hachem,
Ko Hashimoto, Darren A. Plumb, and Mary B. Goldring
22 Isolation and Culture of Human Osteoblasts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 337
Alison Gartland, Robin M.H. Rumney, Jane P. Dillon,
and James A. Gallagher
23 Human Osteoclast Culture and Phenotypic Characterization . . . . . . . . . . . . . . . . . 357
Ankita Agrawal, James A. Gallagher, and Alison Gartland
24 Effects of Temperature Generated from the Holmium: YAG Laser
on Human Osteoblasts in Monolayer Tissue Culture. . . . . . . . . . . . . . . . . . . . . . . . 377
Moustafa I. Hafez, Anne Sandison, Richard R.H. Coombs,
Ian D. McCarthy, and Al-Shymaa M. Hafez
25 Laser Microdissection Microscopy: Application to Cell Culture. . . . . . . . . . . . . . . . 385
Ahlam Mustafa, Cathy Cenayko, Ragai R. Mitry, and Alberto Quaglia
26 Automated Adherent Human Cell Culture (Mesenchymal Stem Cells) . . . . . . . . . . 393
Robert Thomas and Elizabeth Ratcliffe
27 Culturing and Differentiating Human Mesenchymal Stem Cells
for Biocompatible Scaffolds in Regenerative Medicine. . . . . . . . . . . . . . . . . . . . . . . 407
William R. Otto and Catherine E. Sarraf
Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 427