Human Embryonic Stem Cells

✍ Scribed by J. S. Odorico

• Publisher: BIOS Scientific Publ
• Year: 2005
• Tongue: English
• Leaves: 414
• Series: Advanced Methods ( BIOS )
• Edition: 1
• Category: Library

✦ Synopsis

Since the first successful isolation and cultivation of human embryonic stem cells at the University of Wisconsin, Madison in 1998, there has been high levels of both interest and controversy in this area of research.

This book provides a concise overview of an exciting field, covering the characteristics of both human embryonic stem cells and pluripotent stem cells from other human cell lineages. The following chapters describe state-of-the-art differentiation and characterization of specific ectoderm, mesoderm and endoderm-derived lineages from human embryonic stem cells, emphasizing how these can be used to study human developmental mechanisms. A further chapter discusses genetic manipulation of human ES cells. The concluding section covers therapeutic applications of human ES cells, as well as addressing the ethical and legal issues that this research have raised.

✦ Table of Contents

Book Cover......Page 1
Half-Title......Page 2
Title......Page 3
Copyright......Page 4
Table of Contents......Page 5
Contributors......Page 11
Abbreviations......Page 14
Foreword......Page 18
Preface......Page 20
1.2 Derivation and definition of mouse ES cells......Page 23
1.2.2 Self-renewal of ES cells......Page 24
1.2.3 Markers of ES cells......Page 25
1.3.1 Transcriptional regulators......Page 26
1.3.2 LIF signaling......Page 28
1.3.4 LIF signaling via ERKs appears to promote ES cell differentiation......Page 29
1.3.6 Gene profiling of pluripotent cells......Page 31
1.3.7 The ES cell cycle......Page 32
1.4.1 Spontaneous differentiation: the broad potential of ES cells in culture......Page 33
1.4.2 Enrichment for differentiated cell populations......Page 34
1.4.3 Directed differentiation of ES cells in culture......Page 35
1.4.4 Formation of specialized cell types: combining lineage induction with positional information......Page 36
1.4.5 Understanding embryogenesis through ES cell modeling......Page 37
1.5.1 Animal models of development and disease......Page 38
1.5.2 Cell-based therapies and tissue engineering......Page 39
Acknowledgments......Page 40
References......Page 41
2.2.1 Endogenous stem cells......Page 50
2.2.3 Embryonal carcinoma (EC) cells......Page 51
2.2.4 Embryonic stem (ES) cells......Page 54
2.2.5 Embryonic germ (EG) cells......Page 55
2.3 Embryoid body-derived (EBD) cells......Page 58
2.4 Markers of pluripotency......Page 60
References......Page 61
3.2 Stem cells—definition......Page 66
3.4 Adult stem cells—plasticity......Page 67
3.5 Plasticity of hematopoietic bone marrow cells......Page 68
3.6 Plasticity of mesenchymal stem cells......Page 70
3.7 Mulitpotent adult progenitor cells......Page 71
3.8 Plasticity of skeletal muscle cells......Page 72
3.10 Mechanisms of plasticity......Page 73
3.11 Potential uses of adult stem cells......Page 76
References......Page 77
4.1 Introduction......Page 82
4.2 Derivation, growth and morphology of murine and human ES cells......Page 83
4.2.1 Self-renewal factors......Page 84
4.2.2 Novel feeder layers......Page 86
4.2.4 Differentiation in vitro and in vivo......Page 87
4:2.5 Chromosomal alterations......Page 88
4.3.1 Early post-implantation embryonic development and placentogenesis......Page 90
4.3.2 Neurogenesis......Page 91
4.3.4 Hematopoietic development......Page 92
4.3.5 Cardiac development......Page 93
4.4 ES cells: a renewable source of functional cells......Page 94
4.4.2 Mechanistic versus technical differences in mouse and human ES cell differentiation......Page 95
4.4.3 ES cell differentiation as a tool for discovery......Page 96
References......Page 97
5.1.1 Introduction: from bone marrow stromal cells to mesenchymal stem cells......Page 103
5.1.2 A controversial cell system......Page 104
Isolation techniques......Page 105
Standard culture conditions......Page 106
5.1.5 Recent development: MAPCs......Page 107
5.2.1 In vitro differentiation towards mesenchymal lineages......Page 108
5.2.2 Mesenchymal potential assessed through in vivo experiments......Page 110
5.3 Synergy of hMSC and hES research......Page 111
References......Page 113
6.1 Origin and development of the placenla: introduction......Page 120
6.1.1 Primate trophoblast differentiation and placental morphogenesis......Page 121
6.1.2 Trophectoderm formation and trophoblast stem cells: initiating placental development......Page 125
Helix-loop-helix factors......Page 126
Homeobox factors and other factors in trophoblast differentiation......Page 127
6.2 Bridging the mouse-human gap in placental biology......Page 128
6.2.1. Trophoblast differentiation from human ES cells......Page 129
6.2.2 Human ES cells as a model for placental morphogenesis......Page 131
Temtoma formation......Page 132
6.3 Summary and future prospects......Page 133
References......Page 135
7.1 Introduction......Page 139
7.2 Lessons from mouse ES cell-based hematopoiesis......Page 141
7.3 Hematopoiesis from human ES cells: studies to date......Page 142
7.4 Hematopoiesis from human ES cells: the next stage......Page 144
7.5 Human embryonic stem cells, preimplantation genetic diagnosis, and hematopoiesis......Page 147
References......Page 150
8.2.1 Embryonic vasculogenesis......Page 165
8.2.2 Endothelial cell differentiation and vascularization in ES cells......Page 166
Isolation of murine endothelial cells and progenitors......Page 168
8.3.2 Separation using a selectable marker......Page 169
8.4.2 LDL incorporation......Page 170
8.4.4 In vivo vessel formation......Page 171
8.5 Therapeutic applications of endothelial cell progenitors......Page 173
Endothelial cells in other engineered tissue constructs......Page 174
Restenosis......Page 175
8.6 Challenges today and hopes for tomorrow......Page 176
References......Page 177
9.1 Introduction......Page 181
9.2 Neural induction in vertebrates......Page 182
9.3 Embryonic stem cells as a window to mammalian neural development......Page 183
9.4.1 Neural differentiation: common methodology......Page 184
9.4.2 ES cells as a tool to model neural lineage development......Page 185
9.5.1 Neural differentiation from hES cells: modification of mouse protocols......Page 187
9.5.2 Neural specification from hES cells: application of neural induction principles......Page 188
9.5.3 Isolation of neural cells from differentiated human cell population......Page 191
9.5.4 Identity of hES cell-generated neural cells......Page 192
9.5.5 Functional properties of hES cell-derived neurons......Page 193
9.5.6 Engraftability of hES cell-derived neural precursors......Page 194
9.6.2 How to direct hES cells to glia and neurons with regional identities?......Page 195
9.6.4 Will ES cell-derived neural cells be safe for cell therapy?......Page 196
References......Page 197
10.1 Introduction......Page 201
10.2.1 Endoderm formation and pancreatic morphogenesis......Page 202
10.2.2 Endodermal origin of islets......Page 204
10.2.3 Transcription factors involved in pancreas development......Page 205
10.2.4 Patterning of the endoderm and inductive tissue interactions in the early embryo......Page 206
10.2.5 Human pancreas development compared with mice......Page 209
10.3 Islet differentiation from embryonic stem cells......Page 210
10.3.1 Identification of insulin-producing cells......Page 211
10.3.2 Islet lineage differentiation from mouse and human ES cells......Page 213
10.4 Recapitulating developmental pathways of islet differentiation in ES cells......Page 215
10.5 Remaining questions......Page 216
10.6 Summary......Page 217
References......Page 218
11.1 Introduction......Page 226
11.2 Early signals in cardiac development......Page 227
11.3 In vitro differentiation of mouse and human ES cells to cardiomyocytes......Page 228
11.4 Prospects for myocardial regeneration......Page 231
11.4.1 Directing cardiomyocyte differentiation......Page 233
11.4.2 Cardiomyocyte purification......Page 234
11.4.4 In vivo transplantation and development of anti-rejection strategies......Page 235
11.5 Summary......Page 236
References......Page 237
12.1 Introduction......Page 241
12.2.1 Transfection......Page 242
12.2.2 Infection......Page 243
12.2.3 Transient versus stable integration......Page 245
Expression of marker genes in ES cells......Page 247
Over-expression of cellular genes in ES cells......Page 248
Gene targeting in ES cells......Page 249
Silencing gene expression at the post-transcriptional level......Page 250
12.4.1 Genetic engineering for cellular therapy......Page 251
12.4.2 Genetic engineering in nuclear transplantation therapy......Page 252
References......Page 253
13.1 Introduction......Page 257
13.2 Immune profile......Page 258
13.2.2 MHC antigens......Page 259
13.2.4 Alloantigen recognition and the rejection response......Page 261
13.2.5 HLA expression by hES cells and their derivatives......Page 263
13.3.1 HLA matching......Page 264
13.3.2 Oocyte manipulation strategies for immune matching......Page 265
13.3.3 Genetic manipulation of hES cells......Page 267
13.4.1 Immunosuppressive therapy......Page 271
Calcineurin antagonists......Page 272
Sirolimus......Page 273
Side effects of non-specific immunosuppressive therapy......Page 274
13.4.2 Mixed hematopoietic chimerism......Page 275
References......Page 277
14.1 Introduction......Page 283
14.2 Goals for bringing hES cell-based therapy to clinical practice......Page 284
14.2.1 Isolation of well-characterized donor cells in adequate numbers......Page 285
14.2.2 Test functional properties of cells or tissues in vitro......Page 288
14.2.4 Efficacy in animal models of disease......Page 289
14.2.5 Avoidance of immune rejection......Page 291
14.2.6 Safety......Page 293
14.3 Tissue engineering with hES cells......Page 295
14.4.1 Nexus of cellular transplantation and gene therapy for inherited human disorders......Page 296
14.4.2 Continuous local or systemic delivery of bioactive molecules by ex vivo gene therapy using stem cells......Page 297
14.5 Somatic cell nuclear transfer and hES cells......Page 299
14.6 Progress and promise in disease-specific cell therapies......Page 301
14.6.1 Hematologic disorders......Page 302
14.6.2 Neurological diseases......Page 303
14.6.3 Endocrine deficiencies......Page 304
Bone tissue engineering applications......Page 305
14.6 Future......Page 306
References......Page 307
15.1 Introduction......Page 314
15.3.1 hESCs......Page 315
15.3.2 Raw materials......Page 316
15.4 Cell production......Page 317
15.4.1 Development of large-scale production process......Page 318
Undifferentiated hES cells......Page 319
Differentiated cells......Page 320
15.4.2 Cryopreservation and formulation......Page 321
15.4.4 cGMP production......Page 322
References......Page 323
16.1 Federal regulation of embryo research......Page 325
16.2 The origins of the de facto ban on federal funding for embryo research......Page 326
16.3 Origins of the de jure ban on federal funding for embryo research......Page 327
16.4 Origins of the decision to permit general federal funding of research on embryonic stem cell lines......Page 328
16.5 The decision to narrow the eligibility requirements for federal funding of research with human embryonic stem cells......Page 329
16.6.1 Fertilization as the marker of unique personal identity......Page 331
16.6.2 The argument of potentiality......Page 332
16.6.3 Other developmental markers for determining moral status of the embryo or fetus......Page 333
16.6.4 The pluralistic approach to the moral status of the embryo......Page 334
References......Page 336
17.2 Federal statute......Page 338
17.2.1 Funding restrictions imposed by federal agencies......Page 340
17.2.2 The Dickey Amendment, 1996 federal law......Page 341
17.2.3 Guidelines on stem cell research, August 2001......Page 342
17.3 Patent rights, licensing programs and agreements......Page 343
17.3.1 Pertinent patents......Page 345
Academic licensing......Page 346
Commercial licensing strategy......Page 348
17.4 State regulation of research involving embryos......Page 349
17.5 International legal framework......Page 351
17.6 Summary......Page 353
18.1 Introduction......Page 361
18.2 Large-scale isolation of new genes from early embryos and stem cells......Page 362
18.3 Methods for gene expression profiling......Page 363
18.4 Data analysis and bioinformatics......Page 366
18.5 Expression profiling of stem cells......Page 368
18.6 Follow-up study of cDNA microarrays......Page 371
18.7 cDNA microarray analysis of cloned animals......Page 373
18.8 Large-scale functional studies of genes......Page 375
References......Page 376
19.1 Introduction......Page 383
19.2.1 Ionization mechanisms......Page 384
19.2.2 Sample fractionation......Page 385
19.3.1 Mass analyzers......Page 386
19.4.2 Peptide sequencing......Page 388
19.4.3 Proteolysis......Page 389
19.4.5 Post-translational modifications......Page 390
19.4.6 Phosphopeptide enrichment strategies......Page 391
19.5.2 ICAT reagent......Page 392
References......Page 394
United States Government Stem Cell Links......Page 399
Stem Cell Training Programs......Page 400
Society Devoted to Stem Cells......Page 401
Publications Focusing on Stem Cells......Page 402
Index......Page 404