Influence of hepatocyte growth factor and glucose on differentiation of human pancreatic stem cell into insulin-producing cells

Abstract

Objective To investigate the influence factors of expansion and differentiation of human pancreatic stem cells into insulin-producing cells. Methods Human pancreatic stem cells were obtained from induced abortive fetal pancreatic islets. The pancreatic stem cells were expanded in defined inducing-medium for 15 passengers in vivo. The nestin-positive cells and ATP-binding-cassette-superfamily-G2(ABCG2) positive cells in expanded cells were detected by flow cytometer and immunohistochemistry. Expanded pancreatic stem cells were then induced to form islet-like cell clusters (ICCs). Insulin secretion from ICCs was tested by glucose stimulatory insulin secretion (GSIS). Influence of glucose and hepatocyte growth factor on insulin producing ability of ICCs was investigated during pancreatic stem cells differentiation into insulin-producing-cells. Insulin secretion from ICCs that differentiated from 1×105 expanded cells was used to estimate the efficiency of pancreatic stem cells differentiation. Results Nestin-positive cells and ABCG2-positive cells were expanded to 27.9 and 37.8 times when isolated pancreatic stem cells were cultured for 15 passengers. The expanded cells formed ICCs at 15 days in vitro. When hepatocyte growth factor concentration in culture medium was 0, 5, 10, and 15 μg/L, when stimulated with 5.6 mmol/L glucose, insulin secretion from ICCs was 5.00, 12.93, 14.91 and 11.23 μU, respectively. When stimulated with 25.0 mmol/L glucose, insulin secretion was 19.91, 35.53, 47.43 and 23.33 μU, respectively. When glucose concentration in basic culture medium was 0, 2.8, 5.6 and 11.2 mmol/L, insulin secretion was 9.07, 14.21, 6.91 and 3.53 μU, respectively, while stimulated with 5.6 mmol/L glucose. While stimulated with 25.0 mmol/L glucose, insulin secretion was 26.64, 48.97, 37.63 and 34.20 μU. Conclusion Nestin positive cells and ABCG2 positive cells in can be expanded in vitro. Expanded pancreatic stem cells could retain the ability of differentiation into insulin-producing cells and formation of ICCs. A proportionate hepatocyte growth factor and glucose concentration in inductive medium would be in favor of formation of ICCs and insulin secretion during differentiation of pancreatic stem cells into insulin-producing cells. Key words: Stem cells; Differentiation; Insulin