In vitro derivation of functional insulin-producing cells from human embryonic stem cells.
The capacity for self-renewal and differentiation of human embryonic stem (ES) cells makes them a potential source for generation of pancreatic beta cells for treating type I diabetes mellitus. Here, we report a newly developed and effective method, carried out in a serum-free system, which induced human ES cells to differentiate into insulin-producing cells. Activin A was used in the initial stage to induce definitive endoderm differentiation from human ES cells, as detected by the expression of the definitive endoderm markers Sox17 and Brachyury. Further, all-trans retinoic acid (RA) was used to promote pancreatic differentiation, as indicated by the expression of the early pancreatic transcription factors pdx1 and hlxb9. After maturation in DMEM/F12 serum-free medium with bFGF and nicotinamide, the differentiated cells expressed islet specific markers such as C-peptide, insulin, glucagon and glut2. The percentage of C-peptide-positive cells exceeded 15%. The secretion of insulin and C-peptide by these cells corresponded to the variations in glucose levels. When transplanted into renal capsules of Streptozotocin (STZ)-treated nude mice, these differentiated human ES cells survived and maintained the expression of beta cell marker genes, including C-peptide, pdx1, glucokinase, nkx6.1, IAPP, pax6 and Tcf1. Thirty percent of the transplanted nude mice exhibited apparent restoration of stable euglycemia; and the corrected phenotype was sustained for more than six weeks. Our new method provides a promising in vitro differentiation model for studying the mechanisms of human pancreas development and illustrates the potential of using human ES cells for the treatment of type I diabetes mellitus.
Jiang
Wei
W
Department of Cell Biology and Genetics, College of Life Sciences, Peking University, Beijing 100871, China.
Shi
Yan
Y
Zhao
Dongxin
D
Chen
Song
S
Yong
Jun
J
Zhang
Jing
J
Qing
Tingting
T
Sun
Xiaoning
X
Zhang
Peng
P
Ding
Mingxiao
M
Li
Dongsheng
D
Deng
Hongkui
H
D028341:Activins / Q000494:pharmacology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism / Q000628:therapy; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D005326:Fetal Proteins / Q000378:metabolism; D006609:High Mobility Group Proteins / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000096:biosynthesis*; D051379:Mice; D008819:Mice, Nude; D055760:SOXF Transcription Factors; D020825:T-Box Domain Proteins / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D000098908:Brachyury Protein
D028341:Activins; D015415:Biomarkers; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; D005326:Fetal Proteins; D006609:High Mobility Group Proteins; D018398:Homeodomain Proteins; D007328:Insulin; D055760:SOXF Transcription Factors; D020825:T-Box Domain Proteins; D015534:Trans-Activators; D014157:Transcription Factors; D014212:Tretinoin; D000098908:Brachyury Protein; C490003:MNX1 protein, human; C109188:SOX17 protein, human; C422405:activin A; C084010:pancreatic and duodenal homeobox 1 protein