Pancreatic stem cells were isolated and cultured from aborted human fetal pancreases of gestational age 14-20 weeks. They were seeded at a density of 1 × 104 in serum-free media for differentiation into neuron-li...Pancreatic stem cells were isolated and cultured from aborted human fetal pancreases of gestational age 14-20 weeks. They were seeded at a density of 1 × 104 in serum-free media for differentiation into neuron-like cells, expressing β-tubulin III and glial fibrillary acidic protein. These neuron-like cells displayed a synapse-like morphology and appeared to form a neuronal network. Pancreatic stem cells were also seeded at a density of 1 × 105 for differentiation into islet-like cells, expressing insulin and glucagon, with an islet-like morphology. These cells had glucose-stimulated secretion of human insulin and C-peptide. Results suggest that pancreatic stem cells can be differentiated into neuron-like and islet-like cells.展开更多
BACKGROUND: Type 1 diabets is an autoimmune disease caused by the destruction of pancreatic β-cell with an in- creased incidence worldwide in the closing decades of the 20th century. This study was to investigate the...BACKGROUND: Type 1 diabets is an autoimmune disease caused by the destruction of pancreatic β-cell with an in- creased incidence worldwide in the closing decades of the 20th century. This study was to investigate the effects of human umbilical cord serum (UCS) on the proliferation and function of human fetal islet-like cell clusters (ICCs) in vitro. METHODS: Eight fresh pancreatic glands obtained after in- duction of labor with water bag were mildly exposed to col- lagenase V, and the digested cells were cultured in a RPMI- 1640 medium plus 10% pooled UCS or fetal calf serum (FCS) to permit cells attachment and outgrowth of ICCs. RESULTS: In 8 consecutively explanted glands, develop- ment and proliferation of ICCs were observed. In the pre- sence of FCS, the outgrowth of ICC took place on the top of a flbroblast monocellular layer. UCS affected less growth of fibroblasts and increased the formation of ICCs about four-fold compared with explants from the same glands maintained in FCS. In both UCS and FCS, the insulin con- tent of the medium was variable to a certain extent and progressively declined from day 2 to day 6. Dithizone- stained ICCs in UCS suggested that most cell clusters were islet cells ( β-cells), and the purity of islets was estimated 80%-90%. The ultrastructure of the cultured cells showed a large number of granule-containing cells, most of which were identified as β-cells. CONCLUSION: We conclude that in comparison with ex- plants with FCS, the yield of ICCs and purification of islet cells are markedly increased by UCS and may facilitate the proliferation of pancreatic β-cells intended for islet trans- plantation.展开更多
基金supported by the Science and Technology Plan Project of Yantai City (Transplantation of pancreatic islet cells induced from human embryonic stem cells into diabetic animals in vitro), No. 2008142-9
文摘Pancreatic stem cells were isolated and cultured from aborted human fetal pancreases of gestational age 14-20 weeks. They were seeded at a density of 1 × 104 in serum-free media for differentiation into neuron-like cells, expressing β-tubulin III and glial fibrillary acidic protein. These neuron-like cells displayed a synapse-like morphology and appeared to form a neuronal network. Pancreatic stem cells were also seeded at a density of 1 × 105 for differentiation into islet-like cells, expressing insulin and glucagon, with an islet-like morphology. These cells had glucose-stimulated secretion of human insulin and C-peptide. Results suggest that pancreatic stem cells can be differentiated into neuron-like and islet-like cells.
文摘BACKGROUND: Type 1 diabets is an autoimmune disease caused by the destruction of pancreatic β-cell with an in- creased incidence worldwide in the closing decades of the 20th century. This study was to investigate the effects of human umbilical cord serum (UCS) on the proliferation and function of human fetal islet-like cell clusters (ICCs) in vitro. METHODS: Eight fresh pancreatic glands obtained after in- duction of labor with water bag were mildly exposed to col- lagenase V, and the digested cells were cultured in a RPMI- 1640 medium plus 10% pooled UCS or fetal calf serum (FCS) to permit cells attachment and outgrowth of ICCs. RESULTS: In 8 consecutively explanted glands, develop- ment and proliferation of ICCs were observed. In the pre- sence of FCS, the outgrowth of ICC took place on the top of a flbroblast monocellular layer. UCS affected less growth of fibroblasts and increased the formation of ICCs about four-fold compared with explants from the same glands maintained in FCS. In both UCS and FCS, the insulin con- tent of the medium was variable to a certain extent and progressively declined from day 2 to day 6. Dithizone- stained ICCs in UCS suggested that most cell clusters were islet cells ( β-cells), and the purity of islets was estimated 80%-90%. The ultrastructure of the cultured cells showed a large number of granule-containing cells, most of which were identified as β-cells. CONCLUSION: We conclude that in comparison with ex- plants with FCS, the yield of ICCs and purification of islet cells are markedly increased by UCS and may facilitate the proliferation of pancreatic β-cells intended for islet trans- plantation.
