In vivo characterization of transplanted human embryonic stem cell-derived pancreatic endocrine islet cells.
OBJECTIVE
Islet-like clusters (ILCs), differentiated from human embryonic stem cells (hESCs), were characterized both before and after transplantation under the kidney capsule of streptozotocin-induced diabetic immuno-incompetent mice.


METHODS
Multiple independent ILC preparations (n = 8) were characterized by immunohistochemistry, flow cytometry and cell insulin content, with six preparations transplanted into diabetic mice (n = 42), compared to controls, which were transplanted with either a human fibroblast cell line or undifferentiated hESCs (n = 28).


RESULTS
Prior to transplantation, ILCs were immunoreactive for the islet hormones insulin, C-peptide and glucagon, and for the ductal epithelial marker cytokeratin-19. ILCs also had cellular insulin contents similar to or higher than human foetal islets. Expression of islet and pancreas-specific cell markers was maintained for 70 days post-transplantation. The mean survival of recipients was increased by transplanted ILCs as compared to transplanted human fibroblast cells (P < 0.0001), or undifferentiated hESCs (P < 0.042). Graft function was confirmed by secretion of human C-peptide in response to an oral bolus of glucose.


CONCLUSIONS
hESC-derived ILC grafts continued to contain cells that were positive for islet endocrine hormones and were shown to be functional by their ability to secrete human C-peptide. Further enrichment and maturation of ILCs could lead to generation of a sufficient source of insulin-producing cells for transplantation into patients with type 1 diabetes.
Eshpeter
A
A
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Jiang
J
J
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Au
M
M
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Rajotte
R V
RV
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Lu
K
K
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Lebkowski
J S
JS
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Majumdar
A S
AS
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
Korbutt
G S
GS
Alberta Diabetes Institute andDepartment of Surgery, University of Alberta, Edmonton, Canada, andGeron Corporation, Menlo Park, CA, USA.
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D004247:DNA / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000648:ultrastructure; D055098:Endocrine Cells / Q000166:cytology* / Q000648:ultrastructure; D005434:Flow Cytometry; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000648:ultrastructure; D016381:Islets of Langerhans Transplantation; D053208:Kaplan-Meier Estimate; D051379:Mice; D016688:Mice, Inbred NOD
D007328:Insulin; D004247:DNA