Differentiation of embryonic stem cells using pancreatic bud-conditioned medium gives rise to neuroectoderm-derived insulin-secreting cells.
Human embryonic stem cells can be differentiated into insulin-secreting cells by emulating in vitro the key processes that occur during embryonic development. However, the resulting cells are generally immature; thus, further research must be performed to identify the necessary factors to complete the differentiation. To this end, we cultured mouse embryonic stem cells with pancreatic bud-conditioned medium, based on a recent publication. Unlike in humans, mouse cells present two types of insulin, which can be used to identify different cell lineages. As a result, the cell product presented a neuroectodermal genetic expression pattern, with no expression of any definitive endodermal marker analyzed. Also, nonglucose-dependent insulin release was detected. Altogether, this previously published protocol results in neuroectoderm, and not definitive endoderm, derived insulin-positive cells. This further confirms the difficulty of obtaining true cell types of this germ layer. Finally, we identified a 16-kDa protein band that was present in pancreatic bud-conditioned medium. Sequencing this band revealed the presence of Reg proteins. The role of pancreatic bud-conditioned medium remains to be tested in definitive endoderm committed cells.
Research Foundation of Alicante Universitary General Hospital , Hepatology Unit, Alicante, Spain.
Nestor
N
Vicente-Salar
Alfredo
A
Santana
Juan A
JA
Reig
Enrique
E
Roche
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000378:metabolism*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology*; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology / Q000502:physiology*; D051379:Mice; D054258:Neural Plate / Q000166:cytology*; D010179:Pancreas / Q000196:embryology* / Q000378:metabolism*
D017077:Culture Media, Conditioned