葡萄糖浓度与大鼠胰腺导管干细胞的诱导分化

背景：葡萄糖是胰腺导管干细胞分化的重要因素之一,与分化后胰岛素分泌细胞数量及分泌能力相关。目的：对比不同浓度葡萄糖诱导下,胰腺导管干细胞分化后细胞的胰岛素分泌能力。方法：使用胶原酶Ⅴ及Ficoll-400分离及纯化Wistar大鼠胰腺上皮细胞,获取胰腺导管干细胞,将干细胞分为10组,体外培养、增殖及分化形成胰岛素分泌细胞。各组在含有不同浓度葡萄糖的培养基中进行分化。采用免疫荧光染色法鉴定胰腺导管干细胞,光化学发光法检测分化出的胰岛素分泌细胞的胰岛素分泌量。结果与结论：葡萄糖浓度为20.6,25.6,  30.6 mmol/L组细胞的刺激指数高于其他组(P < 0.05),但这3组两两比较差异无显著性意义(P > 0.05)。葡萄糖浓度为15.6,20.6,25.6 mmol/L组细胞胰岛素分泌量高于其他组(P < 0.05),但这3组两两比较差异无显著性意义(P > 0.05)。提示胰腺导管干细胞分化为胰岛素分泌细胞实验中,当分化时培养液所含葡萄糖浓度为20.6~25.6 mmol/L时,所得细胞胰岛素分泌能力最强。

Effects of glucose concentration on differentiation of rat pancreatic duct stem cells

BACKGROUND:Glucose is an important factor on differentiation of pancreatic duct stem cells, it relates to the quantity and secretion function of insulin-producing cells after differentiation.OBJECTIVE: Tocompare the insulin secretion capacity of the differentiated rat pancreatic stem cells induced by various glucose concentrations.METHODS:Rat stem cells were isolated and purified from pancreatic duct cells using collagenase V and Ficoll-400. These stem cells were randomly divided into 10 groups. Every group was induced to culture, proliferate, differentiate and form insulin- producing cells in vitro. The differentiation of all groups was performed in medium with different concentrations of glucose. The immunofluorescence staining was used to identify the pancreatic duct stem cells. The electrochemical luminescence method was used to detect the insulin release from stem cell differentiated islets.RESULTS AND CONCLUSION: The stimulation index of glucose 20.6, 25.6, 30.6 mmol/L groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The insulin releasing of glucose 15.6, 20.6, 25.6 groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The best insulin secretion capacity of insulin-producing cells can be gained by controlling concentration of glucose as 20.6-25.6 mmol/L when pancreatic duct stem cells differentiated into insulin-producing cells.