Characterization of human pancreatic progenitor cells.
β-Cell replacement therapy via islet transplantation is an effective treatment for diabetes mellitus, but its widespread use is severely limited by the shortage of donor organs. Because pancreatic stem/progenitor cells are abundantly available in the pancreas of these patients and in donor organs, the cells could become a useful target for β-cell replacement therapy. We previously established a mouse pancreatic stem cell line without genetic manipulation. In this study, we used the techniques to identify and isolate human pancreatic stem/progenitor cells. The cells from a duct-rich population were cultured in 23 kinds of culture media, based on media for mouse pancreatic stem cells or for human embryonic stem cells. The cells in serum-free media formed "cobblestone" morphologies, similar to a mouse pancreatic stem cell line. On the other hand, the cells in serum-containing medium and the medium for human embryonic stem cells formed "fibroblast-like" morphologies. The cells divided actively until day 30, and the population doubling level (PDL) was 6-10. However, the cells stopped dividing after 30 days in any culture conditions. During the cultures, the nucleus/cytoplasm (N/C) ratio decreased, suggesting that the cells entered senescence. Exendin-4 treatment and transduction of PDX-1 and NeuroD proteins by protein transduction technology into the cells induced insulin and pancreas-related gene expression. Although the duplications of these cells were limited, this approach could provide a potential new source of insulin-producing cells for transplantation.
Noguchi
Hirofumi
H
Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, TX 76104, USA. hirofumn@baylorhealth.edu
Naziruddin
Bashoo
B
Jackson
Andrew
A
Shimoda
Masayuki
M
Ikemoto
Tetsuya
T
Fujita
Yasutaka
Y
Chujo
Daisuke
D
Takita
Morihito
M
Kobayashi
Naoya
N
Onaca
Nicholas
N
Hayashi
Shuji
S
Levy
Marlon F
MF
Matsumoto
Shinichi
S
D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D048430:Cell Shape; D002478:Cells, Cultured; D000077270:Exenatide; D005786:Gene Expression Regulation; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010179:Pancreas / Q000166:cytology*; D010183:Pancreatic Ducts / Q000166:cytology; D010455:Peptides / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic; D014688:Venoms / Q000378:metabolism
D051792:Basic Helix-Loop-Helix Proteins; D000077270:Exenatide; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D010455:Peptides; D015534:Trans-Activators; D014688:Venoms; C084010:pancreatic and duodenal homeobox 1 protein; C093564:Neurogenic differentiation factor 1