표제지
Acknowledgement
목차
Abbreviation 9
Abstract 10
Ⅰ. INTRODUCTION 12
Ⅱ. MATERIALS AND METHODS 15
1. Human ES cell culture 15
2. ES cell cryopreservation by open pulled straw 15
3. ES cell cryopreservation by EM Grids 18
4. Characterization of vitrified ES cells 19
5. In ES vitro cells differentiation and RT-PCR analysis of vitrified 20
6. Statistical Analysis 22
Ⅲ. RESULTS 23
1. Comparison the various vitrification of survival rates protocols under 23
2. Characterization of hES cells, after vitrification 25
Ⅳ. DISCUSSION 27
REFERENCES 40
ABSTRACT IN KOREA 44
Table 1. Schematic procedure on vitrification 30
Table 2. RT-PCR primers used in this study 31
Figure 1. Vitrification vehicles 32
Figure 2. Comparison of survival rates under the various vitrification
conditions 33
Figure 3. Colony forming rates of hES cells frozen-thawed on OPS after
subcultured up to 5th passages 34
Figure 4. Colony forming rates of hES cells frozen-thawed on grids
after subcultured up to 5th passages 35
Figure 5. Colony forming rates after passage down 5 times, following
vitrification supplemented with EG with DMSO and PROH by OPS and
EM grids 36
Figure 6. Immunohistochemistry of vitrified ES cells 37
Figure 7. EB formation and spontaneous differentiation of hES cells 38
Figure 8. Gene expression profiling of vitrified ES cells by differentiation 39