The effect of short-term necrostatin-1 treatment on the differentiation of human induced pluripotent stem beta cells

Abstract

Despite ongoing effort over the past decade, current protocols have been unable to generate mature functional beta cells from human induced pluripotent stem cells (hiPSCs). Based on our previous findings that short-term necrostatin-1 (Nec-1) treatment enhanced pancreatic endocrine cell differentiation and insulin secretion capacity of young porcine islets, we assessed whether short-term treatment of Nec1 can improve the differentiation and function of hiPS beta cells. After 3 days of culture, hiPS beta cells, cultured in either control differentiation media (n=3) or supplemented with Nec-1 (100 μM, n=3), were evaluated for viability, cellular composition, GLUT2 expression in beta cells, and glucose-stimulated insulin expression. While the viability and levels of beta-, alpha-, and GLUT2-positive beta cells were unaffected, the level of insulinand glucagon-positive bi-hormonal cells was significantly lower in Nec-1 treated hiPS beta cells. Short-term Nec-1 treatment also did not affect the insulin secretion ability of hiPS beta cells. The addition of Nec-1 to the differentiation media of hiPS beta cells reduced the number of bi-hormonal cells after short-term culture, suggesting that future studies should evaluate different concentrations and treatment duration of Nec-1.