The reversal of hyperglycaemia in diabetic mice using PLGA scaffolds seeded with islet-like cells derived from human embryonic stem cells.
Islet-like cells derived from embryonic stem (ES) cells may be a promising therapeutic option for future diabetes treatment. Here, we demonstrated a five-stage protocol with adding exendin-4 instead of nicotinamide finally could generate islet-like cells from human embryonic stem (ES) cells. Immunofluorescence analysis revealed a high percentage of c-peptide positive cells in the derivation. However, in addition to insulin/c-peptide, most cells also coexpressed PDX-1 (pancreas duodenum homeobox-1), glucagon, somatostatin or pancreatic polypeptide. Insulin and other pancreatic beta-cell-specific genes were all present in the differentiated cells. Insulin secretion could be detected and increased significantly by adding KCL in high glucose concentration in vitro. Furthermore, subcutaneous transplantation of scaffolds seeded with the islet-like cells or cell transplantation under kidney capsules for further differentiation in vivo could improve 6h fasted blood glucose levels and diabetic phenotypes in streptozotocin-induced diabetic SCID mice. More interestingly, blood vessels of host origin, characterized by mouse CD31 immunostaining, invaded the cell-scaffold complexes. This work reveals a five-stage protocol with adding exendin-4 may be an effective protocol on the differentiation of human ES cells into islet-like cells, and suggests scaffolds can serve as vehicles for islet-like cell transplantation.
Mao
Gen-hong
GH
Reproductive Medical Center, Peking University Third Hospital, Beijing, China.
Chen
Gui-an
GA
Bai
Hai-yan
HY
Song
Tian-ran
TR
Wang
Yan-xia
YX
D000818:Animals; D015415:Biomarkers; D001786:Blood Glucose / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D003921:Diabetes Mellitus, Experimental / Q000150:complications*; D004327:Drinking Behavior / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005215:Fasting / Q000097:blood; D005247:Feeding Behavior / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D006943:Hyperglycemia / Q000097:blood / Q000150:complications* / Q000628:therapy*; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007381:Intermediate Filament Proteins / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D016381:Islets of Langerhans Transplantation; D019344:Lactic Acid / Q000494:pharmacology*; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D011100:Polyglycolic Acid / Q000494:pharmacology*; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D020133:Reverse Transcriptase Polymerase Chain Reaction; D054457:Tissue Scaffolds; D015534:Trans-Activators / Q000378:metabolism
D015415:Biomarkers; D001786:Blood Glucose; D018398:Homeodomain Proteins; D007328:Insulin; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid