Title Page
Contents
Abstract 6
1. Introduction 7
2. Materials & Methods 10
2.1. Materials 10
2.2. Production of Hph-1-PTD and reporter gene plasmids. 10
2.3. Cell culture 12
2.4. In vitro transfection 13
2.5. Flow cytometry 14
2.6. Induction of differentiation 15
3. Results 16
3.1. Production of Hph-1-PTD fusion proteins 16
3.2. Flow cytometry 17
3.3. Adipogenic differentiation 19
3.4. Osteogenic differentiation 22
4. Discussion & Conclusion 26
References 29
Abstract in Korean 34
Figure 1. PTEN working mechanism PTEN is intrinsic lipid phosphatase, which antagonizes the PI3K (phosphoinositide-3-kinase), thereby down-regulating the PI3K-dependent signaling pathways involved... 9
Figure 2. Schematic construction of Hph-1-PTD. 16
Figure 3. Cytofluorimetric identification of ASCs6P and ASCs10P. 18
Figure 4. Oil Red O staining after adipogenic differentiation induction for 2weeks (A) Non-transfected ASCs 6P (B) Non-transfected ASCs 10P (C) PTEN-transfected ASCs 10P Lipid droplet was created and... 20
Figure 5. Quantitative analysis of adipogenic differentiation. Oil Red O dye was destained with 100% isopropanol and optical density was measured at 500nm. (■ Non-transfected 6P ■ Non-transfected 10P □ PTEN-transfected 10P)(이미지참조) 21
Figure 6. Calcein staining after osteogenic differentiation induction for 2weeks. (A) Non-transfected ASCs 6P (B) Non- transfected ASCs 10P (C) PTEN-transfected ASCs 10P. Mineratization was proceeded and... 23
Figure 7. Quantitative analysis of osteogenic differentiation. Alkaline phosphatase activity was gauged by measuring optical density at 500nm. (■ Non-transfected 6P ■Non-transfected 10P □ PTEN -transfected 10P)(이미지참조) 25