Stem Cell and Gene Editing Technology-Based Approaches for Hypoimmunogenic Regenerative Medicine

목차 (Table of Contents)

• ABSTRACT I
• List of Figures IX
• List of Tables XIII
• Abbreviations XIV
• Chapter 1. Manufacturing Hypoimmunogenic iPSC lines and Maintaining Its Immune-modulatory Features via Regulation of Endogenous Antigen Presentation and Processing Machinery genes 1
• 1. Abstract 2
• 2. Introduction 3
• 3. Materials and Methods 6
• 4. Results 21
• 4.1 Generation of human induced pluripotent stem cells 21
• 4.2 Construction of HLA-G-overexpressing K562 cells 27
• 4.3 Ectopic expression of HLA-G suppresses NK cell mediated immune responses 32
• 4.4 Gene editing of hiPSCs using CRISPR/Cas9 34
• 4.5 Pluripotency and genomic stability were not perturbed by gene editing process 40
• 4.6 Cell surface HLA phenotype of differentiated derivatives 47
• 4.7 HLA-G surface expression is perturbed at post-transcriptional level 55
• 4.8 The autophagy pathway is not responsible for HLA-I silencing 58
• 4.9 HLA-I surface presentation and APM gene expression fluctuate during hepatic lineage specification. 60
• 4.10 Downregulation of APM genes restricts HLA-I and transgene expression in NSCs 65
• 4.11 Overexpression of NLRC5 or RelA restored HLA-I cell surface presentation in neural cells 69
• 4.12 Forced expression of RelA recovered HLA-G expression in gene-edited cell lines 73
• 5. Conclusion and discussion 79
• Chapter 2. Impaired CXCR4/SDF-1 Interspecies Interaction Abrogates Endothelial-Blood Cell Adhesion and Chemotactic Migration 84
• 1. Abstract 85
• 2. Introduction 86
• 3. Materials and Methods 90
• 4. Results 98
• 4.1 Comparison in amino acid sequences of CXCR4 and SDF-1 in different mammalian species 98
• 4.2 Porcine CXCR4 is responsive to human SDF-1 104
• 4.3 Cellular responses of PAOEC upon stimulation of CXCR4 by human SDF-1 or porcine SDF-1 109
• 4.4 Porcine SDF-1 partially activates human CXCR4 118
• 4.5 pSDF-1 partially promotes cell proliferation and chemotactic migration in human cells 123
• 4.6 Impaired pSDF-1/CXCR4 signaling abrogates endothelial-blood cell adhesion 132
• 5. Conclusion and discussion 138
• References 142
• Acknowledgement 162