Title Page
Contents
Abstract 13
PART 1. Effect of stem cell transplantation in Q175 Knock-In mice 15
I. INTRODUCTION 15
II. MATERIALS AND METHODS 18
1. Cell culture 18
2. Q175 Knock-In mice and transplantation 18
3. Behavior tests 19
4. Immunohistochemistry 23
5. Western blot analysis 27
6. Statistical analysis 30
III. RESULTS 31
1. Overall experimental procedures 31
2. Survival of transplanted human iPSC-NPCs sorted by CD271-/133+ in 6 months old Q175 Knock-In mice[이미지참조] 34
3. Differentiation of transplanted human iPSC-NPCs sorted by CD271-/133+ in 6 months old Q175 Knock-In mice[이미지참조] 37
4. Weight variance in 6 months old Q175 Knock-In mice transplanted with human iPSC-NPCs sorted by CD271-/133+[이미지참조] 40
5. Behavior test analysis of 6 months old Q175 Knock-In mice transplanted with human iPSC-NPC sorted by CD271-/133+[이미지참조] 43
6. Recovery of neurons degeneration and increase of BDNF in 6 months old Q175 Knock-In mice transplanted with human iPSC-NPCs sorted by CD271-/133+[이미지참조] 54
7. Aggregation of mutant huntington protein decrease in 6 months old Q175 Knock-In mice transplanted human iPSC-NPCs sorted by CD271-/133+[이미지참조] 59
8. Reduced of inflammation in 6 months old Q175 Knock-In mice transplantation iPSC-NPCs sorted by CD271-/133+[이미지참조] 63
IV. CONCLUSION 67
PART 2. Effects of viral vector in Q175 Knock-In mice 70
I. INTRODUCTION 70
II. MATERIALS AND METHODS 72
1. Viral vector 72
2. Q175 knock in mice and injection 72
3. Behavior tests 73
4. Immunohistochemistry 76
5. Statistical analysis 79
III. RESULTS 80
1. Overall experimental method 80
2. Behavior test analysis of 11 months old Q175 Knock-In mice injection viral vector 83
3. Check if the viral vector specifically entered reactive astrocyte 93
4. Reactive astrocyte changes 12 weeks after all behavioral experiments 97
IV. CONCLUSION 102
V. DISCUSSION 104
REFERENCES 106
ABSTRACT IN KOREAN 110
Table 1. List of primary and secondary antibodies for tissue staining. 26
Table 2. List of primary and secondary antibodies for western blot 29
Table 3. List of primary and secondary antibodies for tissue 78
PART I. Effect of cell transplantation in Q175 Knock-In mice 11
Fig. 1. 6 months old Q175 Knock-In mice transplanted with... 33
Fig. 2. Survival of transplanted human iPSC-NPCs sorted by CD271-/133+[이미지참조] 36
Fig. 3. Differentiation of transplanted human iPSC-NPCs sorted by CD271-/133+[이미지참조] 39
Fig. 4. Weight variance of transplanted human iPSC-NPCs... 42
Fig. 5. Motor function behavior test : Rotarod and Grip strength test 46
Fig. 6. Emotional behavior test : Open field test (OFT) 49
Fig. 7. Cognitive behavior test : simple swimming test and Novel... 53
Fig. 8. Observation of recovered in neuron and increased BDNF 58
Fig. 9. Observation of decreased aggregation mHtt protein in... 62
Fig. 10. Reduced the inflammation in Q175 Knock-In mice brain... 66
PART II. Effects of viral vector in Q175 Knock-In mice 12
Fig. 1. 11 months old Q175 Knock-In mice Viral vector injection... 82
Fig. 2. Motor function behavior test : Rotarod and Grip strength test 86
Fig. 3. Emotional behavior test : Open field test (OFT) 89
Fig. 4. Cognitive behavior test : Novel object recognition test 92
Fig. 5. Observation of Viral vector entered 96
Fig. 6. Conversion Reactive astrocyte into Neuron in Dlx2 group 101