Induced differentiation of mouse embryonic stem cells into insulin-secreting cells in vitro / 基础医学与临床

ABSTRACT

Objective To establish a system in which mouse embryonic stem cells (mESCs) differentiate into insulin-secreting cells in vitro. Methods mESCs were cultured to form embryoid bodies(EBs). EBs were cultured in serum-free medium to obtain nestin-positive cells. The selected nestin-positive cells were expanded,then added nicotinamide into the medium to induce the differentiation of nestin-positive cells. Immunochemistry staining and flow cytometry analysis were made on 15th day after induction.Results Flow cytometry analysis revealed that the percentage of nestin-positive cells were different from EBs. The percentage of nestin-positive cells from EBs were higher than those of other diameters. Nestin-positive cells induced by nicotinamide formed islet-like cell clusters. Percentage of insulin-positive cell induced by 10 mmol/L nicotinamide was higher than those induced by 0 mmol/L or 5 mmol/L nicotinamide(P