Title Page
Contents
Abstract 9
I. Introduction 11
II. Materials and methods 14
1) Generation of iPSCs using Sendai virus vector 14
2) iPSC culture 14
3) Vector clearance PCR 15
4) Mycoplasma PCR 15
5) Karyotyping 16
6) Immunocytochemistry 16
7) Cortical neuron differentiation 17
8) ELISA assay 17
9) Cortical organoid differentiation 18
10) Western blot 19
11) Paraffin section 19
12) Section immunocytochemistry 20
13) Statistical analysis 20
III. Results 21
1. Generation and characterization of AD patient-derived iPSCs 21
2. Differentiation of cortical neuron from AD patient-derived iPSCs 25
3. AD pathological analysis in APOE4/4 iPSC-derived neurons 27
4. Differentiation of cortical organoid from AD patient-derived iPSCs 31
5. Amyloid-beta level in APOE4/4 iPSC-derived cortical organoids 34
6. Phospho-tau expression in APOE4/4 iPSC-derived cortical organoids. 37
7. Reduction of Amyloid beta and phosphor-Tau levels by drug treatment in APOE4/4 iPSC-derived cortical organoids 40
IV. Discussion 44
V. Reference 46
Abstract in Korean 50
Figure 1. Virus vector and mycoplasma detection of APOE4/4... 22
Figure 2. Karyotyping analysis of APOE4/4 AD patient-derived iPSC. 23
Figure 3. Pluripotent stem cell marker expression of APOE4/4... 24
Figure 4. Cortical neuron differentiation and characterization of... 26
Figure 5. Increase of Aβ secretion levels in AD patient... 28
Figure 6. Increase of Aβ deposits in AD patient iPSC-derived... 29
Figure 7. Increase of phosphor-tau levels in AD patient... 30
Figure 8. Cortical organoid differentiation from AD... 32
Figure 9. Characterization of AD patient iPSC-derived cortical organoid 33
Figure 10. Increase of intracellular and extracellular Aβ levels in... 35
Figure 11. Increase of Aβ deposits in AD patient iPSC-derived... 36
Figure 12. Increase of phosphor-tau levels in AD patient... 38
Figure 13. Increase of phosphor-tau levels in AD patient... 39
Figure 14. Reduction of Aβ and p-tau levels by drug treatment... 42
Figure 15. Reduction of Aβ and p-tau levels by drug treatment... 43