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Title Page

Abstract

Contents

List of Abbreviations 11

Introduction 12

Materials and Methods 16

Differentiation of photoreceptor precursors from human embryonic stem cells 16

Differentiation of RPEs from human embryonic stem cells 17

Characterization of Photoreceptor Precursors 19

Immunocytochemistry 19

Reverse transcription-polymerase chain reaction (RT-PCR) 19

In vitro phagocytosis assays 21

Animals and Subretinal transplantation of differentiated retinal cells 22

Recording of electroretinogram (ERG) 25

Histology of Retinal Cross-Sections 26

Statistical analysis 27

Results 28

Characterization of differentiated photoreceptor progenitors 28

Characterization of differentiated RPEs 31

Survival and localization of the photoreceptor progenitors after subretinal implantation 35

Retinal precursor cells suppressed ONL thinning in RCS rats 36

Subretinal transplantation of photoreceptor precursors improved ERG in RCS rats 40

Discussion 43

Conclusions 53

References 54

국문초록 59

List of Tables

Table 1. Sequence of primers used in RT-PCR 20

Table 2. Thickness of retinal layers at 4 and 24 weeks after subretinal... 38

Table 3. Amplitude of b-wave in electroretinogram 41

Table 4. Methods for directing differentiation of human pluripotent stem cells to RPE 48

Table 5. Methods for directing differentiation of human pluripotent stem cells to photoreceptor precursors 50

List of Figures

Figure 1. Schematic diagram of differentiation of hESCs into photoreceptor... 18

Figure 2. Immunostaining of the hESC-derived photoreceptor precursors. 29

Figure 3. RT-PCR of the hESC-derived photoreceptor precursors. 30

Figure 4. Confirmation of the hESC-derived RPE cells with expression of the... 32

Figure 5. Images of transmission electron microscope of RPE demonstrate 32

Figure 6. Phagocytosis of the differentiated RPE cells. A confocal microscopic... 33

Figure 7. RT-PCR of the RPE cells differentiated form hESC. 34

Figure 8. Expression of human antigens in the retina 12 weeks after the... 35

Figure 9. Retinal tissue sections stained with hematoxylin and eosin (H&E) at 4... 37

Figure 10. Changes of outer nuclear layer thickness at 4 and 24 weeks after... 39

Figure 11. Changes of b-wave amplitude of ERG at 4, 12 and 24 weeks after... 42

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