title: Generation of pancreatic insulin-producing cells from rhesus monkey induced pluripotent stem cells.
abstract: OBJECTIVE
  The generation of induced pluripotent stem cells (iPSCs) provides a promising possibility for type 1 diabetes therapy. However, the generation of insulin-producing cells from iPSCs and evaluation of their efficacy and safety should be achieved in large animals before clinically applying iPSC-derived cells in humans. Here we try to generate insulin-producing cells from rhesus monkey (RM) iPSCs.
  
  
  METHODS
  Based on the knowledge of embryonic pancreatic development, we developed a four-stage protocol to generate insulin-producing cells from RM iPSCs. We established a quantitative method using flow cytometry to analyse the differentiation efficiency. In addition, to evaluate the differentiation competence and function of RM iPSC-derived cells, transplantation of stage 3 and 4 cells into immunodeficient mice was performed.
  
  
  RESULTS
  RM iPSCs were sequentially induced to definitive endoderm (DE), pancreatic progenitors (PP), endocrine precursors (EP) and insulin-producing cells. PDX1(+) PP cells were obtained efficiently from RM iPSCs (over 85% efficiency). The TGF-β inhibitor SB431542 promoted the generation of NGN3(+) EP cells, which can generate insulin-producing cells in vivo upon transplantation. Finally, after this four-stage differentiation in vitro, insulin-producing cells that could secrete insulin in response to glucose stimulation were obtained. When transplanted into mouse models for diabetes, these insulin-producing cells could decrease blood glucose levels in approximately 50% of the mice.
  
  
  CONCLUSIONS
  We demonstrate for the first time that RM iPSCs can be differentiated into functional insulin-producing cells, which will provide the basis for investigating the efficacy and safety of autologous iPSC-derived insulin-producing cells in a rhesus monkey model for type 1 diabetes therapy.
authors.affiliation: Laboratory of Chemical Genomics, Shenzhen Graduate School of Peking University, Shenzhen, People's Republic of China.
authors.forename: F F
authors.initials: FF
authors.lastname: Zhu
authors.forename: P B
authors.initials: PB
authors.lastname: Zhang
authors.forename: D H
authors.initials: DH
authors.lastname: Zhang
authors.forename: X
authors.initials: X
authors.lastname: Sui
authors.forename: M
authors.initials: M
authors.lastname: Yin
authors.forename: T T
authors.initials: TT
authors.lastname: Xiang
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: M X
authors.initials: MX
authors.lastname: Ding
authors.forename: H
authors.initials: H
authors.lastname: Deng
mesh_terms: D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D064987:Cell- and Tissue-Based Therapy / Q000009:adverse effects; D002478:Cells, Cultured; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000628:therapy*; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy*; D004149:Dioxoles / Q000494:pharmacology; D004195:Disease Models, Animal; D005947:Glucose / Q000494:pharmacology; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007328:Insulin / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism* / Q000637:transplantation; D008253:Macaca mulatta; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D013311:Streptozocin / Q000009:adverse effects; D016896:Treatment Outcome
chemical_list: C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide; D001549:Benzamides; D004149:Dioxoles; D007328:Insulin; D013311:Streptozocin; D005947:Glucose