Establishment of mouse pancreatic stem cell line.
beta-Cell replacement therapy via islet transplantation is a promising possibility for the optimal treatment of type 1 diabetes. However, such an approach is severely limited by the shortage of donor organs. Pancreatic stem/progenitor cells could become a useful target for beta-cell replacement therapy in diabetic patients because the cells are abundantly available in the pancreas of these patients and in donor organs. In this study, we established a mouse pancreatic stem cell line without genetic manipulation. The duct-rich population after islet isolation was inoculated into 96-well plates in limiting dilution. From over 200 clones, 15 clones were able to be cultured for over 3 months. The HN#13 cells, which had the highest expression of insulin mRNA after induction, expressed PDX-1 transcription factor, glucagon-like peptide-1 (GLP-1) receptor, and cytokeratin-19 (duct-like cells). These cells continue to divide actively beyond the population doubling level (PDL) of 300. Exendin-4 treatment and transduction of PDX-1 and NeuroD proteins by protein transduction technology in HN#13 cells induced insulin and pancreas-related gene expression. This cell line could be useful for analyzing pancreatic stem cell differentiation. Moreover, the isolation technique might be useful for identification and isolation of human pancreatic stem/progenitor cells.
Noguchi
Hirofumi
H
Baylor Institute for Immunology Research/Baylor All Saints Medical Center, Baylor Research Institute, Dallas, TX 75204, USA. hirofumn@baylorhealth.edu
Oishi
Koichi
K
Ueda
Michiko
M
Yukawa
Hiroshi
H
Hayashi
Shuji
S
Kobayashi
Naoya
N
Levy
Marlon F
MF
Matusmoto
Shinichi
S
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation; D002469:Cell Separation / Q000379:methods; D000077270:Exenatide; D000067757:Glucagon-Like Peptide-1 Receptor; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology; D007328:Insulin / Q000235:genetics / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000378:metabolism; D053539:Keratin-19 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D010179:Pancreas / Q000166:cytology*; D010183:Pancreatic Ducts / Q000166:cytology; D010455:Peptides / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D018027:Receptors, Glucagon / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014688:Venoms / Q000494:pharmacology
D051792:Basic Helix-Loop-Helix Proteins; D000077270:Exenatide; D000067757:Glucagon-Like Peptide-1 Receptor; D018398:Homeodomain Proteins; D007004:Hypoglycemic Agents; D007328:Insulin; D053539:Keratin-19; D009419:Nerve Tissue Proteins; D010455:Peptides; D012333:RNA, Messenger; D018027:Receptors, Glucagon; D015534:Trans-Activators; D014688:Venoms; C000593833:GLP1R protein, human; C000593834:Glp1r protein, mouse; C084010:pancreatic and duodenal homeobox 1 protein; C093564:Neurogenic differentiation factor 1