Macroencapsulated Human iPSC-Derived Pancreatic Progenitors Protect against STZ-Induced Hyperglycemia in Mice

Abstract

Summary In type 1 diabetes, a renewable source of human pancreatic β cells, in particular from human induced pluripotent stem cell (hiPSC) origin, would greatly benefit cell therapy. Earlier work showed that pancreatic progenitors differentiated from human embryonic stem cells in vitro can further mature to become glucose responsive following macroencapsulation and transplantation in mice. Here we took a similar approach optimizing the generation of pancreatic progenitors from hiPSCs. This work demonstrates that hiPSCs differentiated to pancreatic endoderm in vitro can be efficiently and robustly generated under large-scale conditions. The hiPSC-derived pancreatic endoderm cells (HiPECs) can further differentiate into glucose-responsive islet-like cells following macroencapsulation and in vivo implantation. The HiPECs can protect mice from streptozotocin-induced hyperglycemia and maintain normal glucose homeostasis and equilibrated plasma glucose concentrations at levels similar to the human set point. These results further validate the potential use of hiPSC-derived islet cells for application in clinical settings.

Cell Mass Increase Associated with Formation of Glucose-Controlling β-Cell Mass in Device-Encapsulated Implants of hiPS-Derived Pancreatic Endoderm

“…At end of culture, the preparation contained >90% living cells, the majority with nuclear PDX1 (Pancreatic and Duodenal Homeobox 1) positivity and 50% ± 5% being double‐positive for PDX1 and NKX6.1 (NK6 Homeobox 1) (average for three independent experiments; Supporting Information Fig. S1), similar to the characterization in . An average 10% of cells were single‐ or double‐hormone positive after staining for insulin, glucagon and somatostatin; they were negative for PDX1 and NKX6.1 (Supporting Information Fig.…”

“…The ability of hES‐PE to generate functional β‐cell implants in mice was demonstrated for nonencapsulated as well as for encapsulated grafts . This potential was also observed with human induced pluripotent stem cell‐derived pancreatic endoderm (hiPS‐PE) and with further differentiation stages containing insulin‐positive cells in the grafts .…”

“…To facilitate the investigation of early and late processes in β‐cell formation we wished to identify conditions that help establish PE‐generated β‐cell implants with less variability than in the model studied so far. Our study has been undertaken with hiPS‐PE instead of hES‐PE in our previous work and also applied another device for its encapsulation . The similarity in 20‐week plasma hu‐C‐peptide profiles of comparable cell numbers at start and in variability of individual values supports use of this hiPS‐PE‐device combination to address the aim.…”

“…PE was derived from hiPS cells (produced by Fujifilm Cellular Dynamics, WI) by Nestlé Research (Lausanne, Switzerland) using a scalable and reproducible four‐stage differentiation protocol . Cryopreserved samples were shipped to Brussels.…”

“…Cryopreserved samples were shipped to Brussels. After thawing, aggregates were cultured for 72 hours in “stage 4” medium composed of Dulbecco's modified Eagle's medium high‐glucose with Glutamax, 1% penicillin/streptomycin and 0.5× B27, supplemented by 50 ng/ml Noggin, epidermal growth factor, and keratinocyte growth factor (R&D Systems, MN) and 30 ng/ml Heregulin‐β1 (tebu‐bio, France) . At end of culture, the preparation contained >90% living cells, the majority with nuclear PDX1 (Pancreatic and Duodenal Homeobox 1) positivity and 50% ± 5% being double‐positive for PDX1 and NKX6.1 (NK6 Homeobox 1) (average for three independent experiments; Supporting Information Fig.…”