Title Page
Contents
ABSTRACT 8
Ⅰ. Introduction 10
Ⅱ. Materials and Methods 14
2.1. human iPSC culture and Alkaline Phosphatase (AP) staining 14
2.2. Immunocytochemistry (ICC) 14
2.3. RT-qPCR 15
2.4. Teratoma formation 15
2.5. Karyotyping analysis 15
2.6. Vascular smooth muscle cells differentiation 16
2.7. Generation of Blood vessel organoids 16
2.8. AngioTool analysis 17
2.9. Statistical Analysis 17
Ⅲ. Results 19
3.1. Characterization of control and CADASIL iPSCs 19
3.2. Vascular smooth muscle cell differentiation using control and CADASIL iPSC 22
3.3. Restoring normal phenotype in CADASIL VSMCs by treatment of VEGF and SCF 25
3.4. Generation of blood vessel organoids (BVOs) from CADASIL iPSCs 28
3.5. Analysis of vascular network formation in CADASIL BVOs after growth factor treatment 31
Ⅳ. Discussion 33
References 36
Abstract (in Korean) 39
Table 1. Information about primary antibodies used in Immunocytochemistry. 18
Table 2. Information about primers used in RT-qPCR. 18
Figure 1. Characterization of Human ESCs, iPSC, and CADASIL iPSC 21
Figure 2. Marker expression and NOTCH3 ECD detection in control and CADASIL VSMCs 24
Figure 3. Factor treated control, CADASIL vascular smooth muscle cells (VSMCs) 27
Figure 4. Effect of growth factor treatment on CADASIL iPSC-derived blood vessel organoids (CADASIL BVOs) 30
Figure 5. Analysis of vascular network formation in CADASIL BVOs using AngioTool 32