Title Page
ACKNOWLEGEMENT
Contents
ABBREVIATIONS 10
ABSTRACT 11
1. INTRODUCTION 13
2. MATERIALS AND METHODS 17
2.1. Culture of human embryonic stem cells 17
2.2. Neuronal differentiation of human embryonic stem cell on PA6 18
2.3. Preparation of neurospheres 18
2.4. Expansion of neural precursor cells 19
2.5. Human neural stem cell lines 20
2.6. Culture of ReNcell VM (VM) 21
2.7. Immunocytochemistry 21
2.8. Reverse-transcription Polymerase Chain Reaction (RT-PCR) 22
2.9. Animal model with experimental HD 23
2.10. Preparation of F3.BDNF cells for transplantation 23
2.11. Transplantaion procedures 24
2.12. Rotarod test 24
2.13. Amphetamine-induced rotation test 24
2.14. Modified neurological severity score (mNSS) test 25
2.15. Stepping test 25
2.16. Cresyl violet staining 26
2.17. Immunohistochemistry 26
2.18. Statistical analysis 27
3. RESULTS 28
3.1. Neural differentiation of hESCs on PA6 stromal cells 28
3.2. Morphological characterstics of human neural stem cells used in this study 31
3.3. Expression of neural markers in various cell lines 33
3.4. Expression of human BDNF mRNA and protein 36
3.5. Morphological characteristics of QA-induced HD model 38
3.6. Effects of F3.BDNF cell transplantation 40
3.7. Detection and differentiation of transplanted F3.BDNF cells 42
4. DISCUSSION 44
5. REFERNECES 47
6. SUMMARY (국문요약) 52
Figure 1. Time-course differentiation scheme of human ES cells (CHA-hES3) into neural precursor cells 30
Figure 2. Morphological characteristics of human neural stem cells used in this study 32
Figure 3. Immunocytochemical staining showing the marker expression in various cell lines used in this study 35
Figure 4. Expression of human BDNF mRNA and protein 37
Figure 5. Morphological characteristics of the DA-induced HD rat models 39
Figure 6. Behavioral tests after transplantation 41
Figure 7. Expression of neural and glial markers following transplantation 43