Regulation of osteoclast lineage commitment in bone remodeling

목차 (Table of Contents)

• GENERAL INFORMATION-1
• 1. RESEARCH BACKGROUND-2
• 1-1. Hematopoiesis in bone marrow-2
• 1-2. Bone remodeling-6
• 2. REFERENCES-10
• CHAPTER I. Stem cell factor promotes the expansion of myeloid derived suppressor cells as osteoclast precursors-13
• 1. INTRODUCTION-14
• 2. MATERIALS AND METHODS-20
• 2-1. Antibodies, reagents, cells and mice-20
• 2-2. Protein purification-20
• 2-3. Construction of SCF knock-down 4T1 cells-21
• 2-4. 4T1 induced MDSCs generation-22
• 2-5. MDSCs generation and culture-22
• 2-6. Real-time PCR and immune blotting analysis-23
• 2-7. In vivo mouse cancer models-23
• 2-8. In vivo cytokine injection analysis-24
• 2-9. Osteoclast differentiation and analysis-25
• 2-10. Flow cytometry-26
• 2-11. Statistics-26
• 3. RESULTS-29
• 3-1. SCF derived from metastatic breast carcinoma induce accumulation of MDSCs-29
• 3-2. SCF enhance MDSC accumulation to BM in vivo-35
• 3-3. SCF direct induce MDSC development in presence of GM-CSF in vitro-37
• 3-4. SCF enhances MDSC proliferation and functions-41
• 3-5. SCF promote MDSC derived osteoclast differentiation -45
• 4. DISCUSSION-53
• 5. REFERENCES-57
• CHAPTER II. Pax5 negatively regulates osteoclastogenesis by Blimp1 down-regulation-65
• 1. INTRODUCTION-66
• 2. MATERIALS AND METHODS-71
• 2-1. Antibodies, reagents, cells and mice-71
• 2-2. Plasmid-71
• 2-3. Cell culture and osteoclastogenesis-72
• 2-4. OC analysis-72
• 2-5. Real-time PCR analysis-73
• 2-6. Luciferase reporter assay-74
• 2-7. Generation of transgenic mice-74
• 2-8. Micro-computerized tomography analysis and bone histomorphometry-75
• 2-9. Statistics-75
• 3. RESULTS-79
• 3-1. Pax5 is downregulated during osteoclastogenesis-79
• 3-2. Pax5 negatively regulates RANKL-induced osteoclastogenesis-81
• 3-3. The PD domain of Pax5 is involved in inhibition of RANKL-induced OC differentiation-84
• 3-4. Pax5 reduces Blimp1 activity in osteoclastogenesis -87
• 3-5. Transgenic mice expressing TRAP promoter induced Pax5 to show osteopetrotic phenotype caused by reducing OC formation -90
• 4. DISCUSSION-97
• 5. REFERENCES-100
• SUMMARY (in Korean)-105
• ACKNOWLEDEMENTS-109