Regulation of Apoptosis and Differentiation by p53 in Human ...

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TABLE OF CONTENTS ABBREVIATIONS LIST OF PUBLICATIONS ABSTRACT/RESUMO XII XIV XV/XVII CHAPTER 1 1 Introduction 1.1- HUMAN EMBRYONIC STEM CELLS 3 1.1.1- Criteria to define an embryonic stem cell 5 1.1.2- Stem cell markers 5 1.1.3- Culture methods for hESC propagation: advantages and limitations 7 1.1.4- Maintenance factors of self-renewal and pluripotency 9 1.1.4.1- LIF 9 1.1.4.2- Fibroblast growth factor-2 (FGF-2) 10 1.1.4.3- Transforming growth factor β pathway 10 1.1.4.4- Mouse embryonic fibrobasts secreted factors 11 1.1.4.51- Other factors 11 1.1.5- Strategies to improve clonal survival 12 1.1.6- Cell cycle of hESC 12 1.1.7- Epigenetics 13 1.1.8- hESC signature and gene expression profiling 13 1.1.9- International Stem Cell Initiative: an attempt to compare 14 different hESC cell lines 1.1.10- In vitro Differentiation of hESC 15 1.2- GENETIC STABILITY: HESC AS A MODEL 18 1.3- APOPTOSIS 21 1.3.1- Apoptosis in development and homeostasis 21 1.3.2- Apoptosis in ESC 22 1.3.3- Major molecular regulators of apoptosis 23 1.3.3.1- Bcl2 family 23 1.3.3.2- Caspases 24 1.3.4- Biochemistry of apoptosis 25 1.3.3- Apoptotic pathways 25 1.3.3.1- The Extrinsic Pathway 25 1.3.3.2- The Intrinsic Pathway 25 1.4- TUMOUR SUPPRESSOR P53 27 1.4.1- p53 knockout mice 28 1.4.2- p53 in apoptosis 28 1.4.3- p53 family and differentiation 30 1.4.4- Role of p53 in hESC 31 1.5- RNA INTERFERENCE: SHRNA AND LENTIVIRUS 32 1.5.1- Mechanism of RNAi 32 1.5.2- Tool to study gene function 33 1.6- AIMS AND HYPOTHESIS 35 CHAPTER 2 37 Material and Methods Section 2.1- Cell Culture Methods 40 2.1.1- Commonly Used Tissue Culture Solutions, Reagents and Materials 40 2.1.1.1- Growth factors 40 2.1.1.2- Extracellular matrices 41 2.1.1.3- Enzymes and dissociation solutions 41 2.1.1.4- Miscellaneous Culture Components, Solutions and Material 42 2.1.1.5- Medium solutions 43 - ix -
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Page 10 and 11: 2.1.2- hESC and other cell lines Cu
Page 12 and 13: ABBREVIATIONS Ac- Activin Apaf-1-Ap
Page 14 and 15: LIST OF PUBLICATIONS Article Grande
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Page 20 and 21: CHAPTER 1: Introduction - 2 -
Page 22 and 23: CHAPTER 1: Introduction Trophoectod
Page 24 and 25: CHAPTER 1: Introduction TABLE 1.1-
Page 26 and 27: CHAPTER 1: Introduction Xu et al (X
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CHAPTER 2: Material and Methods Sec
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CHAPTER 2: Material and Methods 8 m
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CHAPTER 2: Material and Methods Con
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CHAPTER 2: Material and Methods fla
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CHAPTER 2: Material and Methods 2.1
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CHAPTER 2: Material and Methods 6.
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CHAPTER 2: Material and Methods Sec
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CHAPTER 2: Material and Methods Rab
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CHAPTER 2: Material and Methods 30%
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CHAPTER 2: Material and Methods TAB
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CHAPTER 3: Characterization of hESC
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CHAPTER 4: RNAi to explore role of
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CHAPTER 5: Conclusions and Future -
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CHAPTER 5: Conclusions and Future W
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CHAPTER 5: Conclusions and Future A
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CHAPTER 5: Conclusions and Future a
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CHAPTER 6: Bibliography - 128 -
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CHAPTER 6: Bibliography Benard, J.,
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CHAPTER 6: Bibliography Duensing, A
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CHAPTER 6: Bibliography Itskovitz-E
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CHAPTER 6: Bibliography Melino, G.,
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CHAPTER 6: Bibliography Rubinson, D
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CHAPTER 6: Bibliography Wei, C. L.,
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CHAPTER 6: Bibliography - 142 -
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APPENDIX I - 144 -
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APPENDIX I 4% Paraformaldehyde (PFA
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TABLE A 2.1- List of genes upregula
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PMP22 1.76 0.70 NM_153322 PDE6G 1.8
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TABLE A 2.2- List of functions of u
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Protein Synthesis Ophthalmic Diseas
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1.73 0.897 CN292254 LOC651029 1.73
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Reproductive System Disease Cellula
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