Title Page
Abstract
Abstract in Korean
Contents
Introduction 14
Material and Method 19
1. Procurement of fetal porcine pancreatic tissue 19
2. Characterization of fetal pancreas in vivo 19
3. Preparation and culture of fetal porcine pancreas 20
4. The removal of mesenchymal cells 20
5. Immunocytochemical analysis 21
6. RNA extraction and reverse transcription PCR assay 22
7. Measurement of insulin secretion 24
8. Xenotransplantation of fetal porcine pancreatic cells to mice 24
9. Morphometry 25
10. Statistical analysis 25
Results 26
1. Transition of fetal porcine and pancreas on various developmental stages 26
2. Cellular events of pancreas on fetal development 28
3. Formation of epithelial monolayer on culture of fetal pancreatic cells 31
4. Nestin expression in fetal porcine pancreatic stem cells 33
5. Differentiation to insulin producing cells from epithelial cells in fetal ductal cell culture 35
6. The self-renewal of fetal pancreatic stem cells 37
7. Morphometric analysis of insulin producing cells and secreted insulin contents in the culture of fetal porcine pancreas 39
8. The role of mesenchymal cells in the proliferation and differentiation of epithelial cells 41
9. Increase of differentiation rate into insulin-producing cells by nicotinamide 45
10. Formation of ductal and islet structure of fetal porcine pancreatic cells in the graft 47
11. Beta cell differentiation of epithelial cells in duct structure 49
12. Transformation of epithelial duct structure to insulin-producing islet 51
Discussion 53
References 58
Figure. 1 Profile of the fetal pig and pancreatic tissues at various stages of development. 27
Figure. 2 Proliferation ability and expression of transcription factors in fetalpancreas at various stages of fetal development. 30
Figure. 3 Culture of the fetal porcine pancreas. 32
Figure. 4 Nestin expression in pancreatic cells prepared from mid-stage. 34
Figure. 5 Transformation of the insulin producing cells from the ductal epithelial cells. 36
Figure. 6 Self-renewal of insulin-producing cells during culture. 38
Figure. 7 Quantification of insulin-producing cell differentiation and insulin secretion. 40
Figure. 8 The formation and expansion of epithelial explants cultured with or without mesenchymal cells. 43
Figure. 9 The effect of the presence of mesenchymal cell for beta cell formation during culture. 44
Figure. 10 Increase of insulin-cell differentiation by nicotinamide. 46
Figure. 11 Formation of cyst and islet structure in graft. 48
Figure. 12 Differentiating epithelial cells on duct-lining in renal graft. 50
Figure. 13 Islet formation of fetal pancreatic stem cells. 52