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首页> 外文期刊>World Journal of Gastroenterology >In vitro cultivation of human fetal pancreatic ductal stem cells and their differentiation into insulin-producing cells
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In vitro cultivation of human fetal pancreatic ductal stem cells and their differentiation into insulin-producing cells

机译：人胎儿胰管干细胞的体外培养及其向胰岛素产生细胞的分化

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AIM: To isolate, culture and identify the human fetal pancreatic ductal stem cells in vitro, and to observe the potency of these multipotential cells differentiation into insulin-producing cells. METHODS: The human fetal pancreas was digested by 1 g/L collagease type Ⅳ and then 2.5 g/L trypsin was used to isolate the pancreatic ductal stem cells, followed by culture in serum-free, glucose-free DMEM media with some additional chemical substrates in vitro (according to the different stage). The cells were induced by glucose-free (control), 5 mmol/L, 17.8 mmol/L and 25 mmol/L glucose, respectively. The cell types of differentiated cells were identified using immunocytochemical staining. RESULTS: The shape of human fetal pancreatic ductal stem cells cultured in vitro was firstly fusiform in the first 2 wk, and became monolayer and cobblestone pattern after another 3 to 4 wk. After induced and differentiated by the glucose of different concentrations for another 1 to 2 wk, the cells formed the pancreatic islet-like structures. The identification and potency of these cells were then identified by using the pancreatic ductal stem cell marker, cytokeratin-19 (CK-19), pancreatic β cell marker, insulin and pancreatic a cell marker, glucagons with immunocytochemical staining. At the end of the second week, 95.2% of the cells were positive for CK-19 immunoreactivity. Up to 22.7% of the cells induced by glucose were positive for insulin immunoreactivity, and less than 3.8% of the cells were positive for glucagon immunoreactivity in pancreatic islet-like structures. The positive ratio of immunoreactive staining was dependent on the concentration of glucose, and it was observed that the 17.8 mmol/L glucose stimulated effectively to produce insulin- and glucagons-producing cells. CONCLUSION: The human fetal pancreatic ductal stem cells are capable of proliferation in vitro. These cells have multidifferentiation potential and can be induced by glucose and differentiated into insulin-producing cells in vitro.
机译：目的：在体外分离，培养和鉴定人胎儿胰管干细胞，并观察这些多能细胞分化为胰岛素产生细胞的潜能。方法：用1 g / LⅣ型胶原酶消化人胎儿胰腺，然后使用2.5 g / L胰蛋白酶分离胰管干细胞，然后在无血清，无葡萄糖的DMEM培养基中与其他化学试剂一起培养体外底物（根据不同阶段）。分别用无葡萄糖（对照），5 mmol / L，17.8 mmol / L和25 mmol / L葡萄糖诱导细胞。使用免疫细胞化学染色鉴定分化细胞的细胞类型。结果：体外培养的人胎儿胰管干细胞的形状在最初的2 wk内首先呈梭状，在随后的3到4 wk内变为单层和鹅卵石型。在由不同浓度的葡萄糖诱导并分化另外1至2周后，细胞形成了胰岛样结构。然后通过使用胰管干细胞标记，细胞角蛋白19（CK-19），胰β细胞标记，胰岛素和胰a细胞标记，胰高血糖素进行免疫细胞化学染色，从而鉴定这些细胞。在第二周结束时，95.2％的细胞对CK-19免疫反应呈阳性。在胰岛样结构中，葡萄糖诱导的细胞中最多有22.7％的胰岛素免疫反应阳性，而胰高血糖素免疫反应的阳性细胞不到3.8％。免疫反应性染色的阳性比例取决于葡萄糖的浓度，并且观察到17.8 mmol / L的葡萄糖可以有效地刺激产生胰岛素和胰高血糖素的细胞。结论：人胎儿胰管干细胞具有体外增殖能力。这些细胞具有多重分化潜能，可以被葡萄糖诱导并在体外分化为胰岛素产生细胞。

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