Induction of insulin-producing cells from human pancreatic progenitor cells.
BACKGROUND
We previously established a mouse pancreatic stem cell line without genetic manipulation. In this study, we sought to identify and isolate human pancreatic stem/progenitor cells. We also tested whether growth factors and protein transduction of pancreatic and duodenal homeobox factor-1 (PDX-1) and BETA2/NeuroD into human pancreatic stem/progenitor cells induced insulin or pancreas-related gene expressions.


METHODS
Human pancreata from brain-dead donors were used for islet isolation with the standard Ricordi technique modified by the Edmonton protocol. The cells from a duct-rich population were cultured in several media, based on those designed for mouse pancreatic or for human embryonic stem cells. To induce cell differentiation, cells were cultured for 2 weeks with exendin-4, nicotinamide, keratinocyte growth factor, PDX-1 protein, or BETA2/NeuroD protein.


RESULTS
The cells in serum-free media showed morphologies similar to a mouse pancreatic stem cell line, while the cells in the medium for human embryonic stem cells formed fibroblast-like morphologies. The nucleus/cytoplasm ratios of the cells in each culture medium decreased during the culture. The cells stopped dividing after 30 days, suggesting that they had entered senescence. The cells treated with induction medium differentiated into insulin-producing cells, expressing pancreas-related genes.


CONCLUSIONS
Duplications of cells from a duct-rich population were limited. Induction therapy with several growth factors and transduction proteins might provide a potential new strategy for induction of transplantable insulin-producing cells.
Noguchi
H
H
Baylor All Saints Medical Center, Baylor Research Institute, Fort Worth, Texas 76104, USA. hirofumn@baylorhealth.edu
Naziruddin
B
B
Shimoda
M
M
Fujita
Y
Y
Chujo
D
D
Takita
M
M
Peng
H
H
Sugimoto
K
K
Itoh
T
T
Tamura
Y
Y
Olsen
G S
GS
Kobayashi
N
N
Onaca
N
N
Hayashi
S
S
Levy
M F
MF
Matsumoto
S
S
D001926:Brain Death; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002467:Cell Nucleus / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure; D004230:Dithizone; D000077270:Exenatide; D051523:Fibroblast Growth Factor 7 / Q000494:pharmacology; D052216:Glucagon-Like Peptide 1 / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D010455:Peptides / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D014019:Tissue Donors; D014688:Venoms / Q000494:pharmacology
D007328:Insulin; D010455:Peptides; D012333:RNA, Messenger; D014688:Venoms; D051523:Fibroblast Growth Factor 7; D004230:Dithizone; D052216:Glucagon-Like Peptide 1; D000077270:Exenatide