Title Page
Contents
ABSTRACT 8
1. Introduction 9
2. Materials and Methods 13
2.1. Mouse embryonic stem cell culture 13
2.2. Differentiation into intermediate mesodermal cells 13
2.3. RNA extraction and real-time RT-qPCR analysis 14
2.4. Immunocytochemistry 14
2.5. Protein extraction and western blotting 14
2.6. Statistical Analysis 15
3. Results 18
3.1. Exploring potential factors for differentiation of mouse embryonic stem cells into intermediate mesodermal cells 18
3.2. Determination of factors for induction of mesodermal differentiation from ESCs 22
3.3. Mesodermal differentiation of mouse embryonic stem cells by treatment of Wnt activator on the later stage of differentiation 26
3.4. Intermediate mesodermal differentiation of mouse embryonic stem cells 28
3.5. Differentiation of intermediate mesodermal cells into müllerian mesoepithelial cells 32
4. Discussion 35
References 38
Abstract (in Korean) 41
Table 1. Primer list for RT-qPCR 16
Table 2. Antibody list for Immunocytochemistry 16
Table 3. Antibody list for Western blot 17
Figure 1. Stepwise signaling pathways in intermediate mesoderm development during mouse embryogenesis.... 12
Figure 2. Induction of intermediate mesodermal lineage from ESCs using Activin, BMP4, CHIR 99021, and Retinoic acid.... 20
Figure 3. Modified culture condition for inducing intermediate mesoderm differentiation from ESCs.... 24
Figure 4. Modified protocol for mesodermal differentiation of mouse ESCs by treatment of Wnt activator on the later stage of differentiation.... 27
Figure 5. Comparative analysis of Wnt activators on the induction of intermediate mesoderm and coelomic epithelium.... 30
Figure 6. Differentiation protocol for the induction of intermediate mesoderm and coelomic epithelium.... 31
Figure 7. Evaluating each subset of Wnt and Hox signaling factors. Quantitative real-time PCR (qRT-PCR) analysis was conducted to examine... 33
Figure 8. Müllerian duct specification using ACBR and RA treatment. Real-time quantitative reverse transcription PCR (qRT-PCR) analysis for assessing the... 34