Ng Yuen Wah. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 118-144). / Abstracts in English and Chinese. / Abstract (English) --- p.ii / (Chinese) --- p.vii / Acknowledgements --- p.x / Publications --- p.xii / Abbreviations --- p.xiv / List of Tables --- p.xix / List of Figures --- p.xx / Table of Contents --- p.xxii / Chapter CHAPTER ONE: --- Introduction --- p.1 / Chapter 1.1 --- Ex Vivo Expansion of Cord Blood CD34+ Cells --- p.1 / Chapter 1.1.1 --- Hematopoiesis --- p.1 / Chapter 1.1.2 --- Hematopoietic Stem Cells --- p.2 / Chapter 1.1.2.1 --- Properties --- p.2 / Chapter 1.1.2.2 --- Functions and Sources of Hematopoietic Stem Cells --- p.3 / Chapter 1.1.2.3 --- Umbilical Cord blood Transplantation --- p.4 / Chapter 1.1.3 --- Ex Vivo Expansion of Hematopoietic Stem Cells --- p.5 / Chapter 1.1.3.1 --- The Need for Ex Vivo Expansion --- p.5 / Chapter 1.1.3.2 --- Ex Vivo Expansion for Clinical Transplantation --- p.6 / Chapter 1.1.3.3 --- Factors Promoting Ex Vivo Expansion of Hematopoietic Stem Cells --- p.7 / Chapter 1.1.3.3.1 --- Cytokines --- p.7 / Chapter 1.1.3.3.2 --- Stromal Feeder Support of Hematopoiesis --- p.9 / Chapter 1.1.3.3.3 --- Regulation of Self-Renewal --- p.11 / Chapter 1.1.4 --- Hematopoiesis and Neurogenesis --- p.12 / Chapter 1.1.4.1 --- Possible Crosstalks --- p.12 / Chapter 1.1.4.2 --- Neurotrophic Factors --- p.13 / Chapter 1.1.4.2.1 --- Brain-Derived Neurotrophic Factor --- p.13 / Chapter 1.1.4.2.2 --- Neurotrophin-3 --- p.14 / Chapter 1.2 --- Embryonic Stem Cells --- p.15 / Chapter 1.2.1 --- Embryonic Stem Cells and Potential Applications --- p.15 / Chapter 1.2.2 --- Maintenance of Undifferentiated State --- p.17 / Chapter 1.2.2.1 --- Mouse Embryonic Stem Cells --- p.17 / Chapter 1.2.2.2 --- Human Embryonic Stem Cells --- p.19 / Chapter 1.2.2.3 --- Embryonic Stem Cell-Specific Markers --- p.20 / Chapter 1.2.3 --- Controlled Differentiation of Embryonic Stem Cells to Hematopoietic Lineages --- p.21 / Chapter 1.2.3.1 --- Formation of Embryoid Body --- p.21 / Chapter 1.2.3.2 --- Hematopoietic Development in Embryoid Body --- p.22 / Chapter 1.2.3.3 --- Factors Promoting Hematopoietic Differentiation --- p.23 / Chapter 1.2.3.3.1 --- Cytokines --- p.23 / Chapter 1.1.3.3.2 --- Stromal Cells --- p.24 / Chapter 1.2.3.3.3 --- Mechanism of Hematopoietic Differentiation --- p.25 / Chapter 1.2.3.4 --- Potential Applications of Embryonic Stem Cells-Derived Hematopoietic Cells --- p.26 / Chapter 1.2.3.5 --- Culture Methods --- p.26 / Chapter 1.2.4 --- Issues Concerning Clinical Applications of Embryonic Stem Cells --- p.28 / Chapter 1.2.4.1 --- Teratoma Formation --- p.28 / Chapter 1.2.4.2 --- Contamination by Mouse Cells --- p.29 / Chapter 1.2.4.3 --- Ethical Issues --- p.29 / Chapter CHAPTER TWO: --- Objectives --- p.31 / Chapter CHAPTER THREE: --- Materials and Methods --- p.33 / Chapter 3.1 --- Ex Vivo Expansion of Cord Blood CD34+ Cells --- p.33 / Chapter 3.1.1 --- Collection of Human Umbilical Cord Blood --- p.33 / Chapter 3.1.2 --- Enrichment of Cord Blood CD34+ Cells --- p.33 / Chapter 3.1.3 --- Ex Vivo Expansion of CD34+ Cells --- p.36 / Chapter 3.1.3.1 --- Establishment of Two Stromal Cell Feeder Layers --- p.36 / Chapter 3.1.3.1.1 --- Primary Mouse AGM Stromal Cells --- p.36 / Chapter 3.1.3.1.2 --- Mouse C17.2 Neural Stem Cell Line --- p.37 / Chapter 3.1.3.2 --- Cryopreservation and Thawing of Stromal Cells --- p.37 / Chapter 3.1.3.3 --- Characterization of AGM --- p.38 / Chapter 3.1.3.4 --- Coculture of Cord Blood CD34+ Cells and Stromal Cells --- p.39 / Chapter 3.1.3.4.1 --- Preparation of Stromal Cells --- p.39 / Chapter 3.1.3.4.2 --- Contact Cocultures of CD34+ Cells and Stromal Cells --- p.39 / Chapter 3.1.3.4.3 --- Effects of Neurotrophic Growth Factors on Ex Vivo Expansion of Enriched CD34+ Cells --- p.40 / Chapter 3.1.4 --- Flow Cytometry Analysis of Human Cell Surface Markers --- p.41 / Chapter 3.1.5 --- Human Progenitor Colony-Forming Unit Assays --- p.42 / Chapter 3.1.6 --- Detection of mRNA Expressions of Hematopoietic and Neurotrophic Growth Factors in Mouse AGM and C17.2 Cells --- p.45 / Chapter 3.1.6.1 --- Cell Culture and Extraction of Total Cellular Ribonucleic Acid (RNA) --- p.45 / Chapter 3.1.6.2 --- Reverse Transcriptase -Polymerase Chain Reaction (RT-PCR) --- p.47 / Chapter 3.1.6.3 --- Gel Electrophoresis and Alkaline Transfer of RT-PCR Products and Southern Blot Analysis and Detection --- p.48 / Chapter 3.1.7 --- Detection of mRNA Expressions of Receptors of Neurotrophic Factors in Enriched CD34+and Expanded Progenitor Cells --- p.51 / Chapter 3.1.7.1 --- Culture of CD34+ Cells --- p.51 / Chapter 3.1.7.2 --- "Extraction of Total Cellular RNA, RT-PCR and Gel Electrophoresis" --- p.51 / Chapter 3.1.7.3 --- DNA Sequencing --- p.52 / Chapter 3.1.8 --- Statistical Analysis --- p.53 / Chapter 3.2 --- In Vitro Differentiation of Mouse Embryonic Stem Cells to Hematopoietic Stem Cells --- p.53 / Chapter 3.2.1 --- Maintenance of Mouse Embryonic Stem Cell --- p.53 / Chapter 3.2.1.1 --- Preparation of Mouse Embryonic Fibroblast Stromal Layer --- p.53 / Chapter 3.2.1.2 --- Culture of Embryonic Stem Cells --- p.55 / Chapter 3.2.2 --- Two-Step In Vitro Differentiation of Embryonic Stem Cells --- p.56 / Chapter 3.2.2.1 --- Collection of Stromal Cells - Conditioned Medium --- p.56 / Chapter 3.2.2.1.1 --- Establishment of Two Stromal Cell Feeder Layers --- p.56 / Chapter 3.2.2.1.2 --- Collection of Conditioned Medium --- p.56 / Chapter 3.2.2.2 --- Primary Differentiation of Embryonic Stem Cell into Embryoid Body --- p.57 / Chapter 3.2.2.2.1 --- Methylcellulose-Based Culture --- p.57 / Chapter 3.2.2.2.2 --- Conditioned Medium-Based Culture --- p.58 / Chapter 3.2.2.3 --- Detection of Hematopoietic Progenitors --- p.59 / Chapter 3.2.3 --- Detection of Embryonic Stem Cell Surface Marker --- p.60 / Chapter CHAPTER FOUR: --- Effects of Novel Microenvironments on the Ex Vivo Expansion of Cord Blood CD34+ Cells --- p.65 / Chapter 4.1 --- Characterization of Primary AGM Stromal Cells --- p.65 / Chapter 4.2 --- Effects of Two Mouse Stromal Cells on Ex Vivo Expansion of CD34+ Cells --- p.65 / Chapter 4.2.1 --- Effects of Primary AGM Cells --- p.66 / Chapter 4.2.2 --- Effects of Neonatal Neural Stem Cell line C17.2 Stromal Cells --- p.66 / Chapter 4.3 --- Expressions of Growth Factors in AGM and C17.2 Cells --- p.67 / Chapter 4.4 --- Effects of Two Neurotrophic Factors on the Clonogenicity of Hematopoietic Progenitor Cells --- p.67 / Chapter 4.4.1 --- Effects of Brain-Derived Neurotrophic Factor --- p.68 / Chapter 4.4.2 --- Effects of Neurotrophin-3 --- p.68 / Chapter 4.5 --- Effects of Two Neurotrophic Factors on the Ex Vivo Expansion of CD34+ Cells --- p.69 / Chapter 4.5.1 --- Effects of Brain-Derived Neurotrophic Factor --- p.69 / Chapter 4.5.2 --- Effects of Neurotrophin-3 --- p.70 / Chapter 4.6 --- Expressions of Receptors of Neurotrophic Factors in Enriched CD34+ Cells and Expanded Progenitor Cells --- p.71 / Chapter 4.6.1 --- mRNA Expressions of BDNF-R(Trk B) and NT-3-R (Trk C) --- p.71 / Chapter 4.6.2 --- DNA Sequencing --- p.72 / Chapter 4.7 --- Discussion --- p.73 / Chapter CHAPTER FIVE: --- Maintenance of Mouse Embryonic Stem Cells and Differentiation to Hematopoietic Lineage in the Presence of AGM and C17.2 Conditioned Media --- p.98 / Chapter 5.1 --- Maintenance of Embryonic Stem Cells --- p.98 / Chapter 5.2 --- Primary (Embryoid Body Formation) and Hematopoietic Differentiation of Embryonic Stem Cells --- p.99 / Chapter 5.2.1 --- Methylcellulose-Based Culture --- p.99 / Chapter 5.2.2 --- Conditioned Medium-Based Culture --- p.100 / Chapter 5.3 --- Discussion --- p.101 / Chapter CHAPTER SIX: --- General Discussion and Conclusion --- p.115 / References
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_325112 |
| Date | January 2005 |
| Contributors | Ng, Yuen Wah., Chinese University of Hong Kong Graduate School. Division of Medical Sciences. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xxv, 144 leaves : ill. (some col.) ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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