Molecular and cellular mechanisms of calcium sensing in CD146+ perivascular cells commitment to osteoblast lineage cells.
Kwok, Po Lam. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. === Includes bibliographical references (leaves 124-130). === Abstracts in English and Chinese. === Thesis/Assessment Committee --- p.i === Abstract --- p.ii === 中文摘要 --- p.v === Acknowledgements --- p.vii === List of Fig...
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| Format: | Others |
| Language: | English Chinese |
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2011
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| Online Access: | http://library.cuhk.edu.hk/record=b5896698 http://repository.lib.cuhk.edu.hk/en/item/cuhk-327507 |
| Summary: | Kwok, Po Lam. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2011. === Includes bibliographical references (leaves 124-130). === Abstracts in English and Chinese. === Thesis/Assessment Committee --- p.i === Abstract --- p.ii === 中文摘要 --- p.v === Acknowledgements --- p.vii === List of Figures --- p.viii === List of Tables --- p.x === Table of Abbreviations --- p.xii === Contents --- p.xix === Chapter Chapter 1 --- General Introduction --- p.1 === Chapter Chapter 2 --- The Biology of Human Umbilical Cord Perivascular Cells (HUCPVs) and Their Potential Applications in Tissue Regeneration === Chapter 2.1 --- INTRODUCTION --- p.5 === Chapter 2.1.1 --- Stem cells --- p.5 === Chapter 2.1.2.1 --- Embryonic stem cells --- p.6 === Chapter 2.1.2.2 --- iPS cells --- p.7 === Chapter 2.1.2.3 --- Somatic stem cells --- p.8 === Chapter 2.1.3 --- Mesenchymal stem cells --- p.9 === Chapter 2.1.4 --- Pericytes --- p.11 === Chapter 2.1.5 --- CD146 positive MSCs --- p.12 === Chapter 2.1.6 --- Human umbilical cord perivascular cells (HUCPVs) --- p.13 === Chapter 2.1.7 --- The biology of stem cell microenvironment (niche) --- p.14 === Chapter 2.1.8 --- Current applications of HUCPVs --- p.17 === Chapter 2.1.9 --- Regenerative medicine --- p.17 === Chapter 2.1.10 --- Applications of stem cells in bone regeneration --- p.19 === Chapter 2.2 --- MATERIALS AND METHODS --- p.22 === Chapter 2.2.1 --- Cell culture --- p.22 === Chapter 2.2.2 --- Preparation of Human Umbilical Cord Perivascular (HUCPV) cells --- p.22 === Chapter 2.2.2.1 --- Isolation of Human Umbilical Cord Perivascular (HUCPV) cells from human umbilical cord --- p.22 === Chapter 2.2.2.2 --- Purification of HUCPV cells --- p.23 === Chapter 2.2.3 --- Immunocytochemsitry --- p.24 === Chapter 2.2.4 --- Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) --- p.25 === Chapter 2.2.4.1 --- Isolation of total cellular RNA --- p.25 === Chapter 2.2.4.2 --- Complementary DNA (cDNA) synthesis --- p.26 === Chapter 2.2.4.3 --- Polymerase chain reaction (PCR) --- p.26 === Chapter 2.2.5 --- Quantitative real-time reverse transcriptionpolymerase chain reaction (qRT-PCR) --- p.30 === Chapter 2.2.6 --- In vitro differentiation assays --- p.33 === Chapter 2.2.6.1 --- Osteogenic differentiation --- p.33 === Chapter 2.2.6.2 --- Adipogenic differentiation --- p.33 === Chapter 2.2.6.3 --- Chondrogenic differentiation --- p.34 === Chapter 2.2.6.4 --- In vitro chondrogenic differentiation on gelfoam® --- p.34 === Chapter 2.2.7 --- Cytochemistry staining --- p.35 === Chapter 2.2.7.1 --- Alkaline Phosphatase staining --- p.35 === Chapter 2.2.7.2 --- Alizarin Red S staining --- p.35 === Chapter 2.2.7.3 --- Oil Red O staining --- p.36 === Chapter 2.2.7.4 --- Alcian Blue staining --- p.36 === Chapter 2.2.8 --- Scanning electron microscopy (SEM) --- p.37 === Chapter 2.2.9 --- Transmission electron microscopy (TEM) --- p.37 === Chapter 2.2.10 --- Paraffin tissue embedding --- p.38 === Chapter 2.2.10 --- Haematoxylin and Eosin staining --- p.38 === Chapter 2.3 --- RESULTS --- p.40 === Chapter 2.3.1 --- Isolation and purification of HUCPVs --- p.40 === Chapter 2.3.2 --- Osteogenic differentiation of HUCPVs under normoxia --- p.41 === Chapter 2.3.3 --- Osteogenic differentiation of HUCPVs under hypoxia --- p.42 === Chapter 2.3.4 --- Adipogenic differentiation of HUCPVs --- p.43 === Chapter 2.3.5 --- Chondrogenic differentiation of HUCPVs --- p.43 === Chapter 2.3.6 --- Chondrogenic differentiation of HUCPVs on gelfoam® --- p.44 === Chapter 2.4 --- DISCUSSION --- p.59 === Chapter Chapter 3 --- Calcium and Calcium-sensing Receptor (CaSR) in osteogenesis === Chapter 3.1 --- INTRODUCTION --- p.62 === Chapter 3.1.1 --- Metabolism of calcium --- p.62 === Chapter 3.1.2 --- Calcium-sensing receptor --- p.64 === Chapter 3.1.2.1 --- The molecular structure of calcium-sensing Receptor (CaSR) --- p.64 === Chapter 3.1.2.2 --- The expression pattern of calciumsensing receptor (CaSR) --- p.67 === Chapter 3.1.2.3 --- The physiological function of calcium-sensing receptor in different tissues or organs --- p.68 === Chapter 3.1.2.4 --- Regulatory role of calcium-sensing receptor in calcium sensing and homeostasis --- p.71 === Chapter 3.1.2.5 --- The role of calcium-sensing receptor in diseases --- p.72 === Chapter 3.1.2.6 --- Genetic animal models targeting calciumsensing receptor --- p.73 === Chapter 3.1.2.7 --- Calcium-sensing receptor in mesenchymal lineage Differentiation --- p.76 === Chapter 3.1.2.8 --- The role of calcium-sensing receptor in the skeleton --- p.76 === Chapter 3.1.3 --- Calcium-sensing receptor related pathway --- p.78 === Chapter 3.1.3.1 --- Cyclic AMP pathway --- p.78 === Chapter 3.1.3.2 --- Cyclic AMP response element-binding protein (CREB) --- p.80 === Chapter 3.2 --- MATERIALS AND METHODS --- p.83 === Chapter 3.2.1 --- Preparation of primary mouse osteoblasts (MOB) from long bone --- p.83 === Chapter 3.2.2 --- Preparation of primary mouse osteoblasts (CMOB) from calvaria --- p.84 === Chapter 3.2.3 --- Immunocytochemistry --- p.84 === Chapter 3.2.4 --- Osteogenic differentiation --- p.85 === Chapter 3.2.3 --- Quantitative real-time reverse transcriptionpolymerase chain reaction (qRT-PCR) --- p.85 === Chapter 3.2.4 --- Cell proliferation measurement by BrdU ELISA (colorimetric) assay --- p.85 === Chapter 3.2.5 --- Western blotting analysis --- p.86 === Chapter 3.2.5.1 --- Preparation of the protein lysate --- p.86 === Chapter 3.2.5.2 --- Protein quantitation --- p.86 === Chapter 3.2.5.3 --- SDS-PAGE --- p.87 === Chapter 3.2.5.4 --- Protein transfer --- p.87 === Chapter 3.2.5.5 --- Immunodetection --- p.88 === Chapter 3.2.6 --- cAMP EIA assay --- p.89 === Chapter 3.3 --- RESULTS --- p.91 === Chapter 3.3.1 --- "Expression of CD 146 and CaSR in HUCPVs, primary mouse long bone osteoblasts and MC3T3-E1 cell line" --- p.91 === Chapter 3.3.2 --- The effect of calcium treatment on the osteogenic differentiation potential of MC3T3-E1 cells under normoxia --- p.91 === Chapter 3.3.3 --- The effect of calcium treatment on the osteogenic differentiation potential of MC3T3-E1 cells under hypoxia --- p.92 === Chapter 3.3.4 --- The effect of calcium treatment on cell proliferation in primary mouse long bone osteoblasts --- p.93 === Chapter 3.3.5 --- The effect of calcium treatment on calcium-sensing receptor expression in primary mouse long bone osteoblasts --- p.94 === Chapter 3.3.6 --- The effect of calcium treatment on calcium-sensing receptor expression in HUCPVs --- p.95 === Chapter 3.3.7 --- The effect of calcium treatment on calcium-sensing receptor expression in primary mouse calvarian osteoblasts --- p.96 === Chapter 3.3.8 --- The effect of calcium treatment on cyclic AMP levels in primary mouse long bone osteoblasts --- p.97 === Chapter 3.4 --- DISCUSSION --- p.117 === Chapter Chapter 4 --- General Discussions --- p.121 === References --- p.124 === Appendices --- p.131 |
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