Wong, Wai Chung. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 113-130). / Abstracts in English and Chinese. / Thesis/Assessment Committee --- p.i / Abstract --- p.ii / 中文摘要 --- p.iv / Acknowledgements --- p.vi / List of Figures --- p.vii / List of Tables --- p.ix / Table of Abbreviations --- p.x / Contents --- p.xv / Chapter Chapter I --- Introduction / Chapter 1.1 --- Stem cells --- p.1 / Chapter 1.1.1 --- Embryonic stem cells --- p.2 / Chapter 1.1.2 --- Adult stem cells --- p.2 / Chapter 1.1.3 --- Artificial stem cells --- p.5 / Chapter 1.2 --- Hair bulge stem cells (HBSC) --- p.7 / Chapter 1.2.1 --- Hair follicle --- p.7 / Chapter 1.2.2 --- The discovery of hair bulge stem cells --- p.9 / Chapter 1.2.3 --- The hair bulge niche --- p.10 / Chapter 1.2.4 --- Molecular markers --- p.12 / Chapter 1.2.5 --- Physiological roles of bulge stem cells --- p.13 / Chapter 1.2.6 --- Pathological roles of bulge stem cells --- p.15 / Chapter 1.2.7 --- Developmental plasticity of bulge stem cells --- p.16 / Chapter 1.3 --- "Regulation of adipogenic, osteogenic and cardiogenic differentiation" --- p.17 / Chapter 1.3.1 --- Regulation of adipogenic differentiation --- p.17 / Chapter 1.3.2 --- Regulation of osteogenic differentiation --- p.18 / Chapter 1.3.3 --- Regulation of cardiogenic differentiation --- p.19 / Chapter 1.4 --- Proteomics --- p.23 / Chapter 1.4.1 --- Definition of proteomics --- p.23 / Chapter 1.4.2 --- Two-dimensional gel electrophoresis (2DGE) --- p.25 / Chapter 1.4.3 --- Mass spectrometry and protein identification --- p.29 / Chapter 1.4.4 --- Other techniques associated with proteomics --- p.34 / Chapter 1.4.5 --- Proteomics and stem cells --- p.36 / Chapter 1.5 --- General summary --- p.37 / Chapter 1.6 --- Aims of my study --- p.38 / Chapter Chapter II --- Materials and Methods / Chapter 2.1 --- Animals --- p.39 / Chapter 2.2 --- Immunohistochemistry --- p.39 / Chapter 2.2.1 --- Histology --- p.39 / Chapter 2.2.2 --- Immunohistochemistry --- p.40 / Chapter 2.3 --- Establishment of hair bulge CD34+ stem cell line --- p.41 / Chapter 2.3.1 --- Isolation of hair bulge explants --- p.41 / Chapter 2.3.2 --- Establishing hair bulge primary cultured cells --- p.42 / Chapter 2.3.3 --- Purification of hair bulge stem cells --- p.43 / Chapter 2.4 --- Karyotyping --- p.45 / Chapter 2.5 --- In vitro differentiation --- p.46 / Chapter 2.5.1 --- Adipogenic differentiation --- p.46 / Chapter 2.5.2 --- Osteogenic differentiation --- p.47 / Chapter 2.5.3 --- Cardiogenic differentiation --- p.48 / Chapter 2.6 --- Proteomic analysis --- p.48 / Chapter 2.6.1 --- Sample preparation --- p.48 / Chapter 2.6.2 --- Quantification of proteins --- p.49 / Chapter 2.6.3 --- First-dimensional separation of proteins 226}0ؤ isoelectric focusing (IEF) --- p.50 / Chapter 2.6.4 --- Second-dimensional separation - sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) --- p.51 / Chapter 2.6.5 --- "Silver staining, imaging and destaining" --- p.52 / Chapter 2.6.6 --- In-gel digestion and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis --- p.53 / Chapter 2.7 --- Histochemistry --- p.54 / Chapter 2.7.1 --- Oil Red O staining --- p.54 / Chapter 2.7.2 --- Alizarin Red S staining --- p.55 / Chapter 2.8 --- Immunocytochemistry --- p.56 / Chapter 2.9 --- Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) --- p.57 / Chapter 2.9.1 --- Isolation of total cellular RNA --- p.57 / Chapter 2.9.2 --- Complementary DNA (cDNA) synthesis --- p.58 / Chapter 2.9.3 --- Polymerase chain reaction and agarose gel electrophoresis --- p.59 / Chapter 2.10 --- Western blot analysis --- p.62 / Chapter 2.10.1 --- Sample preparation and quantification of proteins --- p.62 / Chapter 2.10.2 --- SDS-PAGE --- p.63 / Chapter 2.10.3 --- Protein transfer --- p.64 / Chapter 2.10.4 --- Immunodetection --- p.65 / Chapter 2.11 --- Cell proliferation assay --- p.66 / Chapter 2.11.1 --- Determination of growth pattern --- p.66 / Chapter 2.11.2 --- MTT assay --- p.66 / Chapter 2.12 --- Ultrastructural analysis --- p.67 / Chapter 2.12.1 --- Scanning electron microscopy (SEM) --- p.67 / Chapter 2.12.2 --- Transmission electron microscopy (TEM) --- p.68 / Chapter 2.13 --- Statistical analysis --- p.68 / Chapter Chapter III --- Results / Chapter 3.1 --- Isolation and characterization hair bulge stem cells --- p.69 / Chapter 3.2 --- Directed adipogenic differentiation --- p.70 / Chapter 3.3 --- Directed osteogenic differentiation --- p.71 / Chapter 3.4 --- Ability of cardiogenol C to induce cardiogenesis in HBSCs --- p.71 / Chapter 3.5 --- Comparative proteomic analysis of HBSC cardiogenic differentiation induced by cardiogenol C --- p.72 / Chapter 3.6 --- Role of Wnt signaling pathway in cardiogenol C-induced cardiogenesis --- p.74 / Chapter 3.7 --- Role of chromatin remodeling in cardiogenol C-induced cardiogenesis --- p.75 / Chapter 3.8 --- Legends and tables --- p.76 / Chapter Chapter IV --- Discussion --- p.98 / References --- p.113 / Appendices --- p.131 / Publication --- p.134
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_326511 |
| Date | January 2008 |
| Contributors | Wong, Wai Chung., Chinese University of Hong Kong Graduate School. Division of Anatomy. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, xvii, 134 leaves : ill. ; 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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