Pancreatic Islet-Like Three-Dimensional Aggregates Derived From Human Embryonic Stem Cells Ameliorate Hyperglycemia in Streptozotocin-Induced Diabetic Mice.
We previously reported the in vitro differentiation of human embryonic stem cells (hESCs) into pancreatic endoderm. Here we demonstrate that islet-like three-dimensional (3D) aggregates can be derived from the pancreatic endoderm by optimizing our previous protocol. Sequential treatment with Wnt3a, activin A, and noggin induced a transient upregulation of T and MixL1, followed by increased expression of endodermal genes, including FOXA2, SOX17, and CXCR4. Subsequent treatment with retinoic acid highly upregulated PDX1 expression. We also show that inhibition of sonic hedgehog signaling by bFGF/activin βB and cotreatment with VEGF and FGF7 produced many 3D cellular clusters that express both SOX17 and PDX1. We found for the first time that proteoglycans and vimentin(+) mesenchymal cells were mainly localized in hESC-derived PDX1(+) clusters. Importantly, treatment with chlorate, an inhibitor of proteoglycan sulfation, together with inhibition of Notch signaling significantly increased the expression of Neurog3 and NeuroD1, promoting a transition from PDX1(+) progenitor cells toward mature pancreatic endocrine cells. Purified dithizone(+) 3D aggregates generated by our refined protocol produced pancreatic hormones and released insulin in response to both glucose and pharmacological drugs in vitro. Furthermore, the islet-like 3D aggregates decreased blood glucose levels and continued to exhibit pancreatic features after transplantation into diabetic mice. Generation of islet-like 3D cell aggregates from human pluripotent stem cells may overcome the shortage of cadaveric donor islets for future cases of clinical islet transplantation.
Laboratory of Stem Cells and Tissue Regeneration, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea.
Joong-Hyun
JH
Shim
JongHyun
J
Kim
Jiyou
J
Han
Su Yeon
SY
An
Yu Jin
YJ
Jang
Jeongsang
J
Son
Dong-Hun
DH
Woo
Suel-Kee
SK
Kim
Jong-Hoon
JH
Kim
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000276:immunology / Q000502:physiology; D002460:Cell Line; D003921:Diabetes Mellitus, Experimental / Q000628:therapy*; D004707:Endoderm / Q000166:cytology*; D000066449:Human Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000637:transplantation; D006801:Humans; D006943:Hyperglycemia / Q000276:immunology / Q000628:therapy*; D007515:Islets of Langerhans / Q000166:cytology*; D051379:Mice; D013311:Streptozocin
D013311:Streptozocin