Pancreatic endoderm derived from human embryonic stem cells generates glucose-responsive insulin-secreting cells in vivo

Abstract

Development of a cell therapy for diabetes would be greatly aided by a renewable supply of human beta-cells. Here we show that pancreatic endoderm derived from human embryonic stem (hES) cells efficiently generates glucose-responsive endocrine cells after implantation into mice. Upon glucose stimulation of the implanted mice, human insulin and C-peptide are detected in sera at levels similar to those of mice transplanted with approximately 3,000 human islets. Moreover, the insulin-expressing cells generated after engraftment exhibit many properties of functional beta-cells, including expression of critical beta-cell transcription factors, appropriate processing of proinsulin and the presence of mature endocrine secretory granules. Finally, in a test of therapeutic potential, we demonstrate that implantation of hES cell-derived pancreatic endoderm protects against streptozotocin-induced hyperglycemia. Together, these data provide definitive evidence that hES cells are competent to generate glucose-responsive, insulin-secreting cells.

Pancreas organogenesis: From bud to plexus to gland

“…Furthermore, it is necessary to determine the specific differences in the genetic program between rodents and humans, so that in vitro differentiation is not misled by knowledge gained solely from rodent studies (D'Amour et al, 2006;Kroon et al, 2008). Despite ethical constraints on procurement of human fetal tissue, pioneering studies from Scharfman's group provide prodigious information on human pancreas development.…”

Differentiation of Stem Cells into Insulin-Producing Cells: Current Status and Challenges

“…Two signaling pathways, the Wnt and transforming growth factor (TGF)-b are crucial to induce formation of the definitive endoderm. Differentiation of ESCs into definitive endoderm can be achieved via treatment with activin-A and Wnt3a and confirmed by expression of endodermal markers, including SOX17, FOXA2, GATA4, CXCR4, and cerberus (D'Amour et al 2006;Kroon et al 2008). Nodal, CHIR99021, IDE1 and IDE2 are another molecules that induce development of the definitive endoderm (Borowiak et al 2009;Takenaga et al 2007).…”

“…In the fourth stage (pancreatic endoderm formation), cells are cultured in the absence of all factors except B27 for 3 days. Pancreatic endoderm grafted into immunodeficient mice matures in vivo for several months leading to the generation of glucose responsive insulin secreting cells (Kroon et al 2008). The question is in what way the immature endocrine cells implanted into heterotopic environment, such as the epididymal fat pads, subcutaneous tissue or under kidney capsule differentiate into pancreatic cells.…”

Fish-Esc

“…En effet in vitro seules 14 % des cellules sécrè-tent de l'insuline et elles ne répondent pas au glucose [19]. Les chercheurs de Novocell ont modifié le protocole et encapsulent non pas les cellules différenciées, mais les progéniteurs issus de cellules ESh induites jusqu'au stade d'intestin primitif, laissant ainsi la maturation se poursuivre in vivo, ce qui semble efficace si l'on en juge par la production de peptide C en réponse au glucose [20] ( Figure 4). La duré de vie de ces cellules encapsulées serait de deux ans mais leur capacité de réponse à des variations rapides du glucose reste à démontrer.…”