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Adult mouse intrahepatic biliary epithelial cells induced in vitro to become insulin-producing cells

Nagaya, Masaki ; Katsuta, Hitoshi ; Kaneto, Hideaki ; Bonner-Weir, Susan ; Weir, Gordon C

Journal of endocrinology, 2009-04, Vol.201 (1), p.37-47 [Periódico revisado por pares]

Bristol: BioScientifica

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Título:
Adult mouse intrahepatic biliary epithelial cells induced in vitro to become insulin-producing cells
Autor: Nagaya, Masaki ; Katsuta, Hitoshi ; Kaneto, Hideaki ; Bonner-Weir, Susan ; Weir, Gordon C
Assuntos: Adenoviridae - genetics ; Age Factors ; Animals ; Basic Helix-Loop-Helix Transcription Factors - genetics ; Basic Helix-Loop-Helix Transcription Factors - metabolism ; Bile Ducts, Intrahepatic - cytology ; Bile Ducts, Intrahepatic - metabolism ; Bile Ducts, Intrahepatic - physiology ; Biological and medical sciences ; Cell Transdifferentiation - genetics ; Cells, Cultured ; Endocrine pancreas ; Epithelial Cells - metabolism ; Epithelial Cells - physiology ; Fundamental and applied biological sciences. Psychology ; Gene Expression Profiling ; Herpes Simplex Virus Protein Vmw65 - genetics ; Homeodomain Proteins - genetics ; Homeodomain Proteins - metabolism ; Hormones. Régulation ; Insulin - metabolism ; Insulin-Secreting Cells - metabolism ; Insulin-Secreting Cells - physiology ; Male ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins - genetics ; Nerve Tissue Proteins - metabolism ; Regular papers ; Trans-Activators - genetics ; Trans-Activators - metabolism ; Transduction, Genetic ; Vertebrates: endocrinology
É parte de: Journal of endocrinology, 2009-04, Vol.201 (1), p.37-47
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Descrição: Transdifferentiation of cells from a patient's own liver into pancreatic β-cells could be useful for β-cell replacement. We hypothesized that intrahepatic biliary epithelial cells (IHBECs) could become a new source of insulin-producing cells. IHBECs isolated from adult mice were expanded using our novel culture method termed, collagen-embedded floating culture method (CEFCM). With CEFCM, IHBECs formed three-dimensional ductal cysts and rapidly expanded their number by about 15-fold within 2 weeks. Over 90% of cells were positive for cytokeratin 7 and 19. At day 14, IHBECs were transfected with adenoviral (Ad)- pancreas duodenum homeobox 1 (Pdx-1), NeuroD or Pdx-1/VP16. After 7 additional days in serum- and insulin-free differentiation medium (DM), cell phenotypes were determined by RT-PCR, immunostaining and ELISA for insulin. In DM control IHBECs started to express some endocrine progenitor genes (Neurog3, NeuroD, Nkx6.1, and Pdx-1) but lacked insulin gene (Ins) mRNA. Transduced expression of PDX-1, NEUROD or PDX-1/VP16 led to expression of not only INS but also GLUT2 and prohormone convertase 1 and 2. About 3% of 4000 cells counted in PDX-1/VP16 transduced cultures stained strongly for C-peptide suggesting that a subpopulation may have the capacity for differentiation. Transduced cells released insulin (Ad-PDX-1 0.08±0.05, Ad-NEUROD 0.33±0.09, Ad-PDX-1/VP16 0.37±0.14 ng/1×105 cells after 48 h in culture). IHBECs can be markedly expanded, and then with molecular manipulation a subpopulation of these cells can differentiate towards a β-cell phenotype. This approach may lead to a new source of β-cells that can be used for transplantation in diabetes.
Editor: Bristol: BioScientifica
Idioma: Inglês

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Adult mouse intrahepatic biliary epithelial cells induced in vitro to become insulin-producing cells

Nagaya, Masaki ; Katsuta, Hitoshi ; Kaneto, Hideaki ; Bonner-Weir, Susan ; Weir, Gordon C

Bristol: BioScientifica

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