Masters / Palsson Human Adult Stem Cells

1. Auflage 2009
ISBN: 978-90-481-2269-1
Verlag: Springer Netherlands
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 7, 208 Seiten

Reihe: Human Cell Culture

The aim of volume 7 of Human Cell Culture is to provide clear and precise methods for growing primary cultures of adult stem cells from various human tissues and describe culture conditions in which these adult stem cells differentiate along their respective lineages. The book will be of value to biomedical scientists and of special interest to stem cell biologists and tissue engineers. Each chapter is written by experts actively involved in growing human adult stem cells.

Autoren/Hrsg.

Masters, John

Palsson, Bernhard Ø

Inhaltsverzeichnis

1;Preface;6
2;Contents;7
3;Contributors;9
4;Chapter 1 Neural Progenitors;12
4.1;1.1 Primary Derivation and Selection of hNPC;12
4.1.1;1.1.1 Determination of Fetal Age and Developmental Stage;13
4.1.2;1.1.2 Dissection and Digestion of Human Fetal Periventricular Zone;15
4.1.3;1.1.3 Establishing Primary hNPC Cultures;17
4.2;1.2 Long Term Expansion and Maintenance of hNPC;22
4.2.1;1.2.1 Freezing hNPC Aliquots;23
4.2.2;1.2.2 Thawing Cryopreserved hNPC;24
4.2.3;1.2.3 Establishing Secondary hNPC Cultures;25
4.2.4;1.2.4 General Feeding of hNPC;25
4.2.5;1.2.5 Dissociation of hNPC;27
4.2.6;1.2.6 Replating Dissociated hNPC for General Expansion;31
4.3;1.3 Preservation of Conditioned Medium;31
4.4;1.4 “What About Neurospheres?”;32
4.5;1.5 Characterization of Undifferentiated hNPC;32
4.6;1.6 Functional Diagnostics for Multipotency;34
4.6.1;1.6.1 In Vitro Differentiation;35
4.6.2;1.6.2 In Vivo Differentiation;36
4.7;1.7 Equipment, Supplies, Chemicals, Reagents, and Media;37
4.7.1;1.7.1 Equipment and Consumable Supplies;37
4.7.2;1.7.2 Chemicals and Reagents;37
4.7.3;1.7.3 Media;38
4.7.4;1.7.4 Preparation Notes;38
4.7.5;1.7.5 Normal Plating Volumes;39
4.8;1.8 Introduction to the Tissue Culture (TC) Hood;40
4.8.1;1.8.1 Preparation of the Tissue Culture (TC) Hood;40
4.8.2;1.8.2 Notes on Sterility and Methods of Sterilization;40
4.9;References;42
5;Chapter 2 Multipotent Stromal Cells (hMSCs);56
5.1;2.1 Introduction;56
5.2;2.2 Bone Marrow Procurement and Processing;58
5.3;2.3 Freezing and Recovery of MSCs;63
5.4;2.4 Expansion of Harvested MSCs in Flasks or Cell Culture Chambers (CCCs);65
5.5;2.5 Characterization of MSCs – Colony Forming Unit Assay and Differentiation;72
5.6;2.6 hMSC Surface Epitope Analysis by Flow Cytometry;77
5.7;2.7 Potential Therapeutic Applications;79
5.8;2.8 Summary;80
5.9;References;81
6;Chapter 3 Endothelium;84
6.1;3.1 Introduction and Methodology for Tissue EPC Culture;85
6.2;3.2 Methodology for Isolation of Circulating EPCs;89
6.3;3.3 Protocols for Evaluating the Function of EPCs;91
6.4;3.4 Prospective of Development of EPC Culture System;95
6.5;3.5 Concluding Remarks;97
6.6;References;97
7;Chapter 4 Lung;101
7.1;4.1 Foreword;101
7.2;4.2 Ontogeny, Morphological and Spatial Layers of the Human Lung;102
7.3;4.3 “Stemness” and the Adult Human Lung;106
7.4;4.4 Protocols;110
7.5;References;121
8;Chapter 5 Eye;123
8.1;5.1 Limbal Epithelial Stem Cells Maria Notara, Julie T Daniels;123
8.2;5.2 Culture of Human Corneal Stromal Stem Cells Yiqin Du, James L. Funderburgh;131
8.3;5.3 Neural Differentiation;139
8.4;5.4 Isolation and Culture of M¨ uller Stem Cells from the Adult Neural Retina of the Human Eye G. Astrid Limb;140
8.5;References;147
9;Chapter 6 Colon;153
9.1;6.1 Identification of Stem Cell Markers in Colon Tissues;155
9.2;6.2 Isolation of Colon Stem Cells;159
9.3;6.3 Propagation of Colon Stem Cells;163
9.4;6.4 Differentiation of Cells from Colon Spheres;164
10;Chapter 7 Spermatogonia;167
10.1;7.1 Introduction;167
10.2;7.2 Protocols Developed for Mouse SSC Culture;168
10.3;7.3 Modifications of Mouse Protocols for the Application of SSC Culture from Other Species;174
10.4;7.4 Issues to Be Considered for Development of Human SSC Culture;175
10.5;7.5 Feeder-Free Culture and Bioreactor: Two Techniques that Need to Be Explored in Animal Models for Clinical Application of Human SSC Culture;177
10.6;7.6 Concluding Remarks;178
10.7;References;179
11;Chapter 8 Hair Follicle Pluripotent Stem (hfPS) Cells;181
11.1;8.1 Introduction;182
11.2;8.2 Discussion;185
11.3;8.3 Conclusions;186
11.4;8.4 Materials and Methods;187
11.5;References;190
12;Chapter 9 Pancreas;193
12.1;9.1 Pancreas Development and Function;194
12.2;9.2 Regeneration of ß Cells Occurs Physiologicallyand Pathophysiologically;196
12.3;9.3 Evidence that ß Cells Are Generated from PSC;197
12.4;9.4 Evidence that Islet ß Cells Are Capableof Self-Replication;200
12.5;9.5 The Identity of PSC is Inconclusive;201
12.6;9.6 Future Directions for PSC Research;202
12.7;References;203
13;Chapter 10 Prostate;207
13.1;10.1 Properties of Stem Cells;207
13.2;10.2 Anatomy of the Prostate Gland;208
13.3;10.4 Sources of Human Prostate Tissue;209
13.4;10.5 Tissue Handling;210
13.5;10.6 Tissue Processing;210
13.6;10.7 Identification and Characterization of Putative Prostate Stem Cells;210
13.7;10.8 Cell Sorting;211
13.8;10.9 In Vitro Cell Culture Techniques for Maintenance and Characterization of Stem Cells;211
13.9;10.10 In Vitro Differentiation Potential of Self-Renewing Stem Cells;212
13.10;10.11 Molecular Characterization of Putative Prostate Stem Cells;212
13.11;10.12 Techniques for In Vivo Characterization of Putative Human Prostate Stem Cells;214
14;References;215