Stem Cell Culture (eBook)

Front Cover 1
Stem Cell Culture 4
Copyright Page 5
Contents 6
Contributors 12
Preface 16
Biography 20
Chapter 1: Derivation of Human Embryonic Stem Cells in Standard and Chemically Defined Conditions 22
I. Introduction 22
II. Methods 23
III. Discussion 33
IV. Materials 33
References 34
Chapter 2: Autogeneic Feeders for the Culture of Undifferentiated Human Embryonic Stem Cells in Feeder and Feeder-Free Conditions 36
I. Introduction 37
II. Methods and Materials 38
III. Results 42
IV. Discussion and Summary 46
References 49
Chapter 3: Microporous Membrane Growth Substrates for Embryonic Stem Cell Culture and Differentiation 50
I. Introduction 51
II. Rationale 53
III. Methods 53
IV. Summary 75
References 76
Chapter 4: Culture of Rodent Spermatogonial Stem Cells, Male Germline Stem Cells of the Postnatal Animal 80
I. Introduction 81
II. Rationale 84
III. Methods 88
IV. Materials 99
V. Discussion 101
References 102
Chapter 5: Characterization of Human Amniotic Fluid Stem Cells and Their Pluripotential Capability 106
I. Introduction 107
II. Amniotic Fluid Cell Composition and Stem Cells 108
III. In Vitro Potential of Amniotic Fluid Stem Cells 112
IV. Ex Vivo and In Vivo Application of Amniotic Fluid Stem Cells 115
V. Discussion 118
References 119
Chapter 6: Method to Isolate Mesenchymal-Like Cells from Wharton's Jelly of Umbilical Cord 122
I. Introduction 123
II. Rationale 124
III. Methods 124
IV. Materials 131
V. UCMSCs and Culture Characteristics 136
VI. Discussion 137
References 139
Chapter 7: Isolation, Characterization, and Differentiation of Human Umbilical Cord Perivascular Cells (HUCPVCs) 142
I. Introduction 143
II. Methods 144
III. Concluding Remarks 154
References 155
Chapter 8: Hepatic Stem Cells and Hepatoblasts: Identification, Isolation, and Ex Vivo Maintenance 158
I. Introduction 160
II. Epithelial-Mesenchymal Relationship 160
III. Stem Cells and Lineage Biology 161
IV. Liver Processing 171
V. Fractionation of Liver Cell Subpopulations 183
VI. Feeder Cells 191
VII. Extracellular Matrix 195
VIII. Culture Media 197
IX. Monolayer Cultures of Cells 201
X. Three-Dimensional Systems 206
Appendix 1: Nomenclature, Glossary, and Abbreviations 216
Appendix 2: Details on the Preparation of Buffers/Reagents 228
Appendix 3: Commercial Sources of Reagents 232
Appendix 4: Sources of Primary and Secondary Antibodies for Immunoselection and for Characterization of the Cells 233
References 234
Chapter 9: Culture of Pluripotent Neural Epithelial Progenitor Cells from E9 Rat Embryo 248
I. Introduction 249
II. Rationale 250
III. Methods 250
IV. Materials 258
V. Discussion 258
VI. Summary 259
References 260
Chapter 10: Primary and Multipassage Culture of Human Fetal Kidney Epithelial Progenitor Cells 262
I. Introduction 263
II. Rationale 264
III. Methods 266
IV. Materials 271
V. Discussion 274
VI. Summary 275
References 275
Chapter 11: Isolation of Human Mesenchymal Stem Cells from Bone and Adipose Tissue 278
I. Introduction 279
II. Isolation of hMSCs from Trabecular Bone 280
III. Isolation of hMSCs from Adipose Tissue 284
IV. Growth, Subculture, and Cryopreservation of Undifferentiated hMSCs 286
V. Surface Marker Characterization of hMSCs 288
VI. hMSC Differentiation Assays for Osteogenic, Adipogenic, and Chondrogenic Pathways 291
VII. Summary 297
References 297
Chapter 12: Culture of Mesenchymal Stem/Progenitor Cells in Adhesion-Independent Conditions 300
I. Introduction 301
II. Suspension Culture of Bone Marrow-Derived MPCs 301
III. Analysis of Suspension Culture 304
IV. Mechanism of MPC Expansion in Suspension 309
V. Discussion 312
References 313
Chapter 13: Purification and Long-Term Culture of Multipotent Progenitor Cells Affiliated with the Walls of Human Blood Vessels: Myoendothelial Cells and Pericytes 316
I. Introduction 318
II. Materials and Methods: Cell Analysis and Purification by Flow Cytometry 319
III. Materials and Methods: Long-Term Culture of Human Blood Vessel-Associated Progenitor Cells 322
IV. Materials and Methods: Proliferation Kinetics of Human Myoendothelial Cells and Pericytes 324
V. Materials and Methods: Genotypic and Phenotypic Analyses of Long-Term Cultured Cells 325
VI. Conclusion 327
References 329
Chapter 14: Isolation and Culture of Colon Cancer Stem Cells 332
I. Introduction 333
II. Identification of CSCs in Colon Cancer Tissue 335
III. Isolation and Propagation of Colon CSCs 336
IV. Discussion 343
References 344
Chapter 15: Isolation and Establishment of Human Tumor Stem Cells 346
I. Introduction 347
II. Rationale 348
III. Methods 348
IV. Results 357
V. Discussion 359
VI. Materials 361
References 362
Chapter 16: Stem Cells from Cartilaginous and Bony Fish 364
I. Introduction to Models and Uses 365
II. Embryonal Stem Cells 369
III. Tissue-Specific Stem Cells 374
IV. Outlook and Future Contributions 382
References 383
Index 390
Volumes In Series 400