Growth inhibitors promote differentiation of insulin-producing tissue from embryonic stem cells

Abstract

The use of embryonic stem cells for cell-replacement therapy in diseases like diabetes mellitus requires methods to control the development of multipotent cells. We report that treatment of mouse embryonic stem cells with inhibitors of phosphoinositide 3-kinase, an essential intracellular signaling regulator, produced cells that resembled pancreatic ␤ cells in several ways. These cells aggregated in structures similar, but not identical, to pancreatic islets of Langerhans, produced insulin at levels far greater than previously reported, and displayed glucose-dependent insulin release in vitro. Transplantation of these cell aggregates increased circulating insulin levels, reduced weight loss, improved glycemic control, and completely rescued survival in mice with diabetes mellitus. Graft removal resulted in rapid relapse and death. Graft analysis revealed that transplanted insulin-producing cells remained differentiated, enlarged, and did not form detectable tumors. These results provide evidence that embryonic stem cells can serve as the source of insulin-producing replacement tissue in an experimental model of diabetes mellitus. Strategies for producing cells that can replace islet functions described here can be adapted for similar uses with human cells.

Directed differentiation of human embryonic stem cells towards a pancreatic cell fate

“…It has been shown that pancreatic cell types can be generated from mouse and non-human primate ESCs [10][11][12][13][14][15][16][17][18]. Based on these findings, recent studies have also suggested the potential of hESCs to differentiate into insulin-producing cells through spontaneous in vitro differentiation [5] or the use of a multi-stage protocol [6].…”

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

“…Boyd et al (2008) compared three different protocols to differentiate ESCs into IPCs using the embryoid body formation strategy. This study indicated that the IPCs generated using Blyszczuk protocol (Blyszczuk et al 2004) exhibited superior C-peptide expression and longer normoglycemic rescue in diabetic mice when compared with the Hori's and Lumelsky's protocols (Hori et al 2002;Lumelsky et al 2001). In protocol described by Blyszczuk the ESCs were cultured in ''handing drops'' and in suspension for 5 days to form embryoid bodies.…”

“…The Lumelsky's protocol involves serum-free ITSFn medium and basic fibroblast growth factor (bFGF) treatments (Lumelsky et al 2001). The Hori's protocol involves additionally treatment of differentiating cells with PI3 kinase inhibitor LY294002, but excludes B27 supplement (Hori et al 2002). Real-time polymerase chain reaction and immunofluorescence techniques indicated that each of these protocols lead to differentiation of ESCs toward pancreatic lineage.…”

“…The potential of IPCs generated from ESCs using embryoid body approach to normalize glucose in streptozotocin-induced diabetes mice is very low. Several reports indicated transient correction of blood glucose level after IPCs transplantation (Blyszczuk et al 2003;Hori et al 2002;Lumelsky et al 2001). This may be due to cell dying or dedifferentiation of transplanted IPCs.…”

“…This may be due to cell dying or dedifferentiation of transplanted IPCs. Another problem regarding the ESCs differentiation into IPCs using embryoid body strategy is teratoma formation (Blyszczuk et al 2003;Hori et al 2002;Boyd et al 2008). A possible explanation for such a finding may be that IPCs are in fact a heterogenous mixture of undifferentiated ESCs, IPCs and even differentiated cells of other lineages.…”