Title page
CONTENTS
1. Introduction 12
2. Materialsandmethods 17
2.1. CultureofhumanES cells 17
2.2. DifferentiationofhumanES cellsintoneuralprecursorcells 17
2.2.1. Formationofembryoidbody 17
2.2.2 Derivationandisolationofneuralprecursorcells 18
2.3. Differentiationofneuralprecursorcellsintoneuronandglialcells 18
2.3.1. Differentiaionofneuralprecursorcellsintoneurons 18
2.3.2. Differentiationofneuralprecursorcellsintogliallineages 19
2.4. Preparationsofneuralprecursorcellsforanalyses 19
2.4.1. Cryosectionofneuralprecursorcells 19
2.4.2. Sectionofneuralprecursorcellsinparaffin 19
2.5. Histochemicalandimmunocytochemicalanalyses 20
2.5.1. Histochemicalanalyses 20
2.5.2. Immunocytochemicalanalysis 20
2.6. RT-PCR analyses 20
3. Results 24
3.1. CharacterizationofhumanES cells 24
3.2. DifferentiationhumanES cellsintoneuralprecursorcells 24
3.2.1. FormationofEmbryoidbodies 24
3.2.2. DifferentiationofEBsintoneuralprecursorcells 27
3.3. Characterizationoftheneuralprecursorcells 27
3.3.1. ConfirmationofNeuralprecursorcells 27
3.3.2. Differentiationcapacityofneuralprecursorcells 29
4. Discussion 36
5. Conclusions 40
6. References 41
7. Summary in korean 48
Table 1. Antibodiesusedintheimmunocytochemicalanalyses 22
Table 2. PrimersusedinRT-PCR analyses 23
Figure 1. Characterization of cellular markers for human ES cells 25
Figure 2. Morphological assessments during differentiation of human ES cells into neural precursor cells and neurons in vitro 26
Figure 3. RT-PCR and immunocytochemistry analyses in EBs 28
Figure 4. RT-PCR analyses for neural precursor cells specific marker in neural precursor cells 30
Figure 5. Immunocytochemical analyses for cellular markers specific for neural precursor cells(scale bar 100㎛) 31
Figure 6. RT-PCR analyses of neuron-specific markers in neural precursor cells and their differentiated neurons 33
Figure 7. Immunocytochemical analyses of neuron-specific markers in neurons differentiation from neural precursor cell(scale bar 100㎛) 34
Figure 8. Immunocytochemical analyses of glial markers in cells differentiated from neural precursor cells(scale bar 100㎛) 35