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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20.6,25.6,30.6mmol/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15.6,20.6,25.6mmol/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20.6~25.6mmol/L时,所得细胞胰岛素分泌能力最强。
BACKGROUND: Glucose is one of the important factors in the differentiation of pancreatic ductal stem cells, which is related to the number of insulin-secreting cells and the secretory capacity after differentiation. OBJECTIVE: To compare the insulin secretion ability of pancreatic ductal stem cells under the induction of different concentrations of glucose. Methods: Pancreatic ductal epithelial cells of Wistar rats were isolated and purified by collagenase Ⅴ and Ficoll-400. Pancreatic ductal stem cells were obtained. The stem cells were divided into 10 groups and cultured, proliferated and differentiated to form insulin-secreting cells. Groups were differentiated in medium containing different concentrations of glucose. Pancreatic ductal stem cells were identified by immunofluorescence staining, and the amount of insulin secreted by the differentiated insulin-secreting cells was measured by photochemical luminescence. RESULTS AND CONCLUSION: The stimulation index of cells with glucose concentration of 20.6, 25.6 and 30.6 mmol / L was higher than that of other groups (P <0.05). However, there was no significant difference between the three groups (P> 0.05). The insulin secretion of cells with glucose concentration of 15.6, 20.6 and 25.6 mmol / L was higher than that of other groups (P <0.05), but there was no significant difference between the three groups (P> 0.05). Tip pancreatic ductal stem cells into insulin-secreting cells experiment, when the culture medium containing glucose concentration of 20.6 ~ 25.6mmol / L, the resulting cell insulin secretion ability strongest.