title page
Acknowledgement
Contents
Abbreviation 10
Abstract 12
I. Introduction 13
II. Materials and methods 15
II.1. Preparation of lentiviral vectors 15
II.2. Mice 16
II.3. HSC purification and viral transduction 16
II.4. Colony-forming-cell capacity assay 17
II.5. Direct intra-bone marrow (IBM) injection of lentiviral vector 17
II.6. FACS analysis 18
III. Results 19
III.1. Lentiviral vector constructs 19
III.2. Retronectin enhancement of lentiviral gene transfer into HSC/HPCs with multilineage potential in vitro. 21
III.3. Effects of Retronectin for gene delivery into HSC/HPCs in vivo. 25
IV. Discussion 32
V. Conclusion 37
VI. References 38
Summary in Korean 45
Table 1. Transgene marking in lymphoid and myleoid as well as HSCs in bone marrow 1 week after IBM injection of RN and LV. 29
Table 2. Transgene distribution in Hematopoietic and non-hematopoietic system of mice 1 week after IBM injection with RN and LV. 31
Figure 1. Lentiviral vector constructs 20
Figure 2. Retronectin enhancement of gene transfer into c-Kit+ Lin-BM cells in vitro by FACS.(이미지참조) 23
Figure 3. Effects of Retronectin on efficiency of gene transfer into immature subsets of CFU-C with multilineage capabilities in vitro. 24
Figure 4. Retronectin augments of gene transduction of c-Kit+ Lin-BM cells from femur and tibia of mice by FACS.(이미지참조) 28
Figure 5. Effects of Retronectin on efficiency of gene transduction of high proliferative CFU-C with multilineage capabilities in vivo. 30