Human induced pluripotent stem cells differentiate into insulin-producing cells able to engraft in vivo.
OBJECTIVE
New sources of insulin-secreting cells are strongly required for the cure of diabetes. Recent successes in differentiating embryonic stem cells, in combination with the discovery that it is possible to derive human induced pluripotent stem cells (iPSCs) from somatic cells, have raised the possibility that patient-specific beta cells might be derived from patients through cell reprogramming and differentiation. In this study, we aimed to obtain insulin-producing cells from human iPSCs and test their ability to secrete insulin in vivo.


METHODS
Human iPSCs, derived from both fetal and adult fibroblasts, were differentiated in vitro into pancreas-committed cells and then transplanted into immunodeficient mice at two different stages of differentiation (posterior foregut and endocrine cells).


RESULTS
IPSCs were shown to differentiate in insulin-producing cells in vitro, following the stages of pancreatic organogenesis. At the end of the differentiation, the production of INSULIN mRNA was highly increased and 5 ± 2.9 % of the cell population became insulin-positive. Terminally differentiated cells also produced C-peptide in vitro in both basal and stimulated conditions. In vivo, mice transplanted with pancreatic cells secreted human C-peptide in response to glucose stimulus, but transplanted cells were observed to lose insulin secretion capacity during the time. At histological evaluation, the grafts resulted to be composed of a mixed population of cells containing mature pancreatic cells, but also pluripotent and some neuronal cells.


CONCLUSIONS
These data overall suggest that human iPSCs have the potential to generate insulin-producing cells and that these differentiated cells can engraft and secrete insulin in vivo.
Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy.
Silvia
S
Pellegrini
Stem Cells and Neurogenesis Unit, Division of Neuroscience, IRCCS San Raffaele Scientific Institute, 20132, Milan, Italy.
Federica
F
Ungaro
Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy.
Alessia
A
Mercalli
Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy.
Raffaella
R
Melzi
Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, 53100, Siena, Italy.|Fondazione Umberto Di Mario ONLUS, c/o Toscana Life Sciences, 53100, Siena, Italy.
Guido
G
Sebastiani
Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, 53100, Siena, Italy.|Fondazione Umberto Di Mario ONLUS, c/o Toscana Life Sciences, 53100, Siena, Italy.
Francesco
F
Dotta
Stem Cells and Neurogenesis Unit, Division of Neuroscience, IRCCS San Raffaele Scientific Institute, 20132, Milan, Italy.
Vania
V
Broccoli
Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy. piemonti.lorenzo@hsr.it.
Lorenzo
L
Piemonti
Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Via Olgettina 60, 20132, Milan, Italy. sordi.valeria@hsr.it.
Valeria
V
Sordi
D000818:Animals; D002096:C-Peptide / Q000032:analysis / Q000378:metabolism; D002454:Cell Differentiation; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts; D005470:Fluorometry; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D007328:Insulin / Q000032:analysis / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000502:physiology* / Q000637:transplantation*; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016035:Pancreas Transplantation
D002096:C-Peptide; D007328:Insulin
Diabetes; Differentiation; Induced pluripotent stem cells; Insulin-producing cells