Title Page
Contents
ABSTRACT 9
Ⅰ. Introduction 10
Ⅱ. Material and Method 12
2.1. Derivation and culture of porcine fetal fibroblast (PFF) 12
2.2. Reprogramming PFFs by Sendai-virus 12
2.3. LCDMV cell culture 12
2.4. Alkaline phosphatase (AP) staining 13
2.5. Embryoid body (EB) formation and In-vitro three-germ layer differentiation 13
2.6. Immunocytochemistry 14
2.7. RNA isolation and real-time RT-qPCR 14
2.8. Electron microscopy 15
2.9. Mitochondrial length measurement 15
2.10. Oxygen consumption rate analysis (OCR) 15
2.11. Extracellular acidification rate analysis (ECAR) 16
2.12. Statistical analysis 16
Ⅲ. Results 18
3.1. Generation and Characterization of Porcine Extended Pluripotent Stem Cells 18
3.2. Differentiation potential of pEPSC into embryonic and extraembryonic lineage 21
3.3. Change of Mitochondrial Morphology during Reprogramming 23
3.4. Mitochondrial Metabolism Profiles in pEPSCs 25
Ⅳ. Discussion 28
References 31
Abstract (in Korean) 35
Table 1. Antibodies used for immunocytochemistry 17
Table 2. Primer sets used for real-time qPCR 17
Figure 1. Generation and characterization of porcine extended pluripotent stem cells 20
Figure 2. Differentiation of pEPSC into embryonic and extraembryonic lineages 22
Figure 3. Analysis of Mitochondrial Morphology in PFFs and pEPSCs 24
Figure 4. Mitochondrial Respiratory Function Analysis in PFFs and pEPSC 26
Figure 5. (A) Measurement of extracellular acidification rate (ECAR) in PFFs and pEPSC. (B) Basal glycolysis (C) Compensatory glycolysis (D) % of Proton efflux... 27