Title Page

Contents

Abstract 11

Ⅰ. INTRODUCTION 13

Ⅱ. MATERIALS AND METHODS 16

1. Human iPSC culture 16

2. Differentiation into three germ layers from iPSCs 17

3. Differentiation into striatal medium spiny neurons 17

4. Differentiation into neuronal precursor cell 19

5. Flow cytometry 19

6. Karyotyping Test 19

7. Immunocytochemistry 20

8. Western blot analysis 22

9. Statistical analysis 23

Ⅲ. RESULTS 24

1. Cell seeding number optimization of HLA-homozygous iPSC 24

2. Characterization of iPSC with the same HLA-haplotype 26

3. Neural precursor cell differentiation and drug screening 31

4. Modeling of medium spiny neuron and characterization 34

Ⅳ. DISCUSSION 44

Ⅴ. CONCLUSION 48

REFERENCES 49

ABSTRACT IN KOREAN 54

Table 1. List of primary and secondary antibodies for cell staining 21

Table 2. List of primary and secondary antibodies for western blot 23

Table 3. Analysis used to characterization iPSC in this study 30

Fig. 1. Characterization of cells per area of HLA #1-iPSC 25

Fig. 2. Characterization of iPSC with the most frequent HLA haplotype in the Korean population 28

Fig. 3. Characterization of iPSC with the second frequent HLA haplotype in the Japanese population 30

Fig. 4. Characterization of NPC and detection of aggregated huntingtin protein at HD patient iPSC-derived NPC. 32

Fig. 5. Reduction of pathological markers by HDAC 1/3 inhibitor in HD patient iPSC-derived NPC. 33

Fig. 6. Differentiation into MSNs refer to the Meng Li protocol 36

Fig. 7. Representative images of immunofluorescence for particular stage markers during medium spiny neuron differentiation. DARPP32,... 37

Fig. 8. Comparison of medium spiny neuron specific marker level by week of differentiated neuron 38

Fig. 9. Representative images of immunofluorescence for particular stage markers during GABAergic neuron differentiation. MAP2,... 39

Fig. 10. Comparison of GABAergic neuron specific marker level by week of differentiated neuron 41

Fig. 11. Comparison of actrocyte marker level by week of differentiated neuron. 42

Fig. 12. Comparison of the pathological marker EM48 of each cell at 12 weeks of differentiation 43