Development of Versatile Drug Delivery Systems for Cell Therapy

목차 (Table of Contents)

• Abstract 1
• Chapter 1: Introduction 5
• 1.1 Overview of cell therapy 6
• 1.2 Advantages of cell-based therapeutics 10
• 1.3 History and recent trends and development of cell therapy 12
• 1.4 Clinical benefits and applications of cell therapy 15
• 1.4.1 Clinical islet cell-based therapy 15
• 1.4.2 Clinical stem cell-based therapy 19
• 1.4.3 Cell-based therapeutics for the treatment of cancer 23
• 1.4.4 Cell-based based therapeutics for the treatment of immune disorders 26
• 1.4.5 Cell therapy in regenerative medicine 27
• 1.5 Limitations of cell therapy 29
• 1.6 Need for effective drug delivery system in therapy 33
• 1.7 Research rationale and drug delivery system for cell therapy 34
• Chapter 2: Prolonged immunomodulatory drug-releasing microspheres and molecular shielding of islet cells via PEGylation for treatment of type 1 diabetic mellitus 38
• 2.1 Introduction 39
• 2.2 Methods and materials 45
• 2.2.1 Donors and Recipients 45
• 2.2.2 Pancreatic islet isolation and culture 46
• 2.2.3 Surface modification of islet cells via PEGylation 46
• 2.2.4 Study of islet cell viability 47
• 2.2.5 Study of islet cell functionality 49
• 2.2.6 Fabrication and characterization of RM-Ms 50
• 2.2.7 Induction of diabetes and transplantation of islet cells 51
• 2.2.8 Intraperitoneal glucose tolerance test 51
• 2.2.9 Isolation of immune cells from spleen, PLN, graft and GAT 51
• 2.2.10 Generation of bone marrow-derived dendritic cells 52
• 2.2.11 Flow cytometry 52
• 2.2.12 RNA isolation and quantitative real-time PCR 53
• 2.2.13 Statistical analysis 54
• 2.3 Results 54
• 2.3.1 Viability and functionality study of PEGylated islet cells 55
• 2.3.2 Transplantation of PEGylated islet cells into subcutaneous space 58
• 2.3.3 In vitro study of activation of costimulatory markers of APCs 60
• 2.3.4 In vivo immunological study 61
• 2.3.5 PCR analysis for the study of the expression of cytokine levels 63
• 2.3.6 Characterization of rapamycin-releasing microspheres 65
• 2.3.7 In vitro study of activation of DCs under rapamycin releasing microspheres 67
• 2.3.8 Co-delivery of PEGylated islet cells and rapamycin-releasing microspheres in the subcutaneous space 70
• 2.4 Discussions 73
• 2.5 Conclusion 81
• Chapter 3: Curcumin-laden ECM-mimicking microfibers assemble with mesenchymal stem cells to generate hybrid spheroids and enhance cell viability and function 83
• 3.1 Introduction 84
• 3.2 Materials and methods 87
• 3.2.1 Preparation of microfibers 88
• 3.2.2 Preparation of fragmented microfibers 88
• 3.2.3 Polydopamine and collagen-I conjugation on the fragmented fibers 89
• 3.2.4 Characterization of fragmented fibers 89
• 3.2.4.1 Solid state characterization 89
• 3.2.4.2 Quantification of collagen-I conjugation on FFs 90
• 3.2.4.3 Drug release profile 90
• 3.2.5 Cell culture 91
• 3.2.6 Preparation of spheroids and heterospheroids 91
• 3.2.7 Assessment of cell viability 91
• 3.2.8 Preparation of cell extracts and western blot analysis 93
• 3.2.9 Statistical analysis 94
• 3.3 Results 94
• 3.3.1 Characterization of fibers 94
• 3.3.2 Characterization of curcumin-loaded fragmented fibers 98
• 3.3.3 Cell viability of spheroids and heterospheroids 101
• 3.3.4 Assessment of cell viability with curcumin-loaded fibers 103
• 3.4 Discussion 106
• 3.5 Conclusion 110
• Chapter 4: Catechol-modified stabilizer facilitates impulsive ligand functionalization of particulates and membrane modification of cells 112
• 4.1 Introduction 113
• 4.2 Materials and methods 114
• 4.2.1 Synthesis and characterization of D-PEMA 114
• 4.2.2 Preparation and characterization of NPs 116
• 4.2.3 Conjugation of ligands and quantification 116
• 4.2.3.1 Ovalbumin conjugation and quantification 117
• 4.2.3.2 DEC205 conjugation and quantification 118
• 4.2.3.3 Galactosamine conjugation and quantification 117
• 4.2.3.4 Folate conjugation and quantification 118
• 4.2.4 Evaluation of nanoparticle toxicity 118
• 4.2.5 Cellular uptake studies 119
• 4.2.6 Surface modification of pancreatic islets 120
• 4.2.7 Statistical analysis 122
• 4.3 Results 122
• 4.3.1 Synthesis and characterization of D-PEMA 122
• 4.3.2 Formulation of D-PEMA NPs 126
• 4.3.3 Effect of D-PEMA NPs on cell viability 128
• 4.3.4 Conjugation of biological ligands on the surface of D-PEMA NPs 130
• 4.3.5 Cellular uptake study of ligand conjugated NPs 131
• 4.3.6 Conjugation of D-PEMA NPs on the surface of islet cells 134
• 4.4 Discussion 135
• 4.5 Conclusion 142
• References 144
• Abstract in Korean 175