Document Detail

Adult mouse intrahepatic biliary epithelial cells induced in vitro to become insulin-producing cells.
MedLine Citation:
PMID:  19168505     Owner:  NLM     Status:  MEDLINE    
Transdifferentiation of cells from a patient's own liver into pancreatic beta-cells could be useful for beta-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/1x10(5) 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 beta-cell phenotype. This approach may lead to a new source of beta-cells that can be used for transplantation in diabetes.
Masaki Nagaya; Hitoshi Katsuta; Hideaki Kaneto; Susan Bonner-Weir; Gordon C Weir
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-01-23
Journal Detail:
Title:  The Journal of endocrinology     Volume:  201     ISSN:  1479-6805     ISO Abbreviation:  J. Endocrinol.     Publication Date:  2009 Apr 
Date Detail:
Created Date:  2009-03-26     Completed Date:  2009-07-23     Revised Date:  2011-06-01    
Medline Journal Info:
Nlm Unique ID:  0375363     Medline TA:  J Endocrinol     Country:  England    
Other Details:
Languages:  eng     Pagination:  37-47     Citation Subset:  IM    
Research Division, Section on Islet Transplantation and Cell Biology, Joslin Diabetes Center, One Joslin Place, Boston, Massachusetts 02215, USA.
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MeSH Terms
Adenoviridae / genetics
Age Factors
Basic Helix-Loop-Helix Transcription Factors / genetics,  metabolism
Bile Ducts, Intrahepatic / cytology,  metabolism,  physiology*
Cell Transdifferentiation / genetics*
Cells, Cultured
Epithelial Cells / metabolism,  physiology*
Gene Expression Profiling
Herpes Simplex Virus Protein Vmw65 / genetics
Homeodomain Proteins / genetics,  metabolism
Insulin / metabolism
Insulin-Secreting Cells / metabolism,  physiology*
Mice, Inbred C57BL
Nerve Tissue Proteins / genetics,  metabolism
Trans-Activators / genetics,  metabolism
Transduction, Genetic*
Reg. No./Substance:
0/Basic Helix-Loop-Helix Transcription Factors; 0/Herpes Simplex Virus Protein Vmw65; 0/Homeodomain Proteins; 0/Nerve Tissue Proteins; 0/Trans-Activators; 0/pancreatic and duodenal homeobox 1 protein; 11061-68-0/Insulin; 169238-82-8/NeuroD protein

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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