| 20236034 |
Mason MN, Mahoney MJ: Inhibition of gamma-secretase activity promotes differentiation of embryonic pancreatic precursor cells into functional islet-like clusters in PEG hydrogel culture. Tissue Eng Part A. 2010 Mar 18.
We assessed the ability of a gamma-secretase inhibitor to promote the in vitro differentiation of induced embryonic pancreatic precursor cell aggregates into functional islet-like clusters when encapsulated within a three-dimensional hydrogel. Undifferentiated pancreatic precursor cells were isolated from E.15 rat embryos, dissociated into single cells, and aggregated in suspension-rotation culture. Aggregates were photoencapsulated into poly (ethylene glycol) hydrogels with entrapped collagen type 1 and cultured for 14 days with, and without, a gamma-secretase inhibitor. Gene expression, proinsulin content, and C-peptide release were measured to determine differentiation and maturation of encapsulated precursor cell aggregates. In control media, scattered breakthrough beta cell differentiation was observed; however, cells remained largely insulin negative. Upon addition of a gamma-secretase inhibitor the majority of cells in clusters became insulin positive and insulin per DNA and glucose stimulated insulin release measurements for these cultures were comparable those for adult rat islets. Cluster counts after culture day 14 were 88% of those initially encapsulated, demonstrating excellent cluster survival in hydrogel culture. These results indicate that concerted differentiation of pancreatic precursor cell aggregates into functionally mature, islet-like clusters can be achieved in PEG-based hydrogel cultures by blocking cell contact-mediated Notch signaling with a gamma-secretase inhibitor. |
35(0,1,1,5) |