Title Page

Contents

Abstract 9

Ⅰ. INTRODUCTION 11

Ⅱ. MATERIALS AND METHODS 15

2.1. HEK293 cell culture 15

2.2. Treated microtubule stabilizer and disturber 15

2.3. OCR measurement 16

2.4. Immunocytochemistry assay 17

2.5. Mitochondrial function analysis 17

2.6. Gene expression analysis 18

2.7. Western blot assay 19

2.8. Embryo collection 20

2.9. Embryo culture and treatment stabilizer, disturber 21

2.10. Blastocyst embryo single cell PCR, Real-time PCR analysis 22

2.11. Statistical Analysis 23

Ⅲ. RESULTS 24

3.1. Microtubule stabilizer affect to morphology, viability, and proliferation of HEK293 24

3.2. Microtubule stabilizer increases mitochondrial oxygen consumption and ATP production 25

3.3. Microtubule stabilizer affects to mitochondria dynamic properties 25

3.4. Microtubule stabilizer activates mTOR and AMPK, a critical regulator of mitochondrial metabolic activity 26

3.5. Microtubule stabilizer increase embryo development 27

3.6. Microtubule stabilizer activates Oct 3/4, Nanog, and SOX2 28

Ⅳ. DISCUSSION 37

Ⅴ. CONCLUSION 41

REFERENCES 42

ABSTRACT IN KOREAN 45

FIGURE 1. Microscope images, cell size, confocal images, cell viability, and cell proliferation of microtubule stabilizers- and disturber-treated cells. (A) Light... 29

FIGURE 2. Analysis of mitochondrial respiration in microtubule stabilizer and disturber treated cells. (A) Mitochondrial respiration, indicated by the OCR, in (a) Taxol-treated (b)... 30

FIGURE 3. Confocal live imaging to analyze mitochondrial mass and mitochondrial membrane potential in microtubule stabilizer and disturber treated cells. (A) Control and... 31

FIGURE 4. Gene expression analysis of MFN1, MFN2, OPA1, and DRP1 in microtubule stabilizer and microtubule disturber treated cells. (A) Single cell PCR... 32

FIGURE 5. Protein expression of AMPK and mTOR in microtubule stabilizer and disturber treated cells. (A) Blots of AMPK, pAMPK, mTOR, and pmTOR in... 33

FIGURE 6. Development time and number of embryos in each stage (4cell, 8cell, morula, and blastocyst) with microtubule stabilizer, disturber treated and control... 34

FIGURE 7. Hatched out blastocyst embryo with microtubule stabilizer and disturber treated. (A) Graph of percentage of hatched out blastocyst. (B) Graph of hatched... 35

FIGURE 8. (A) Single cell PCR analysis of Oct 3/4, Nanog, and SOX2 in microtubule stabilizer-treated blastocyst and disturber-treated blastocyst. (B~D)... 36