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将人单克隆胰腺干细胞(1.0×104个/孔)接种在铺有明胶的6孔板内,体外定向诱导其分化为包含大量β细胞的类胰岛。双硫腙染色呈阳性。RT-PCR检测,转录表达胰岛素。Sprague-Dawley成年大鼠30只,6只腹腔注射缓冲液作正常对照,24只腹腔注射2%链脲菌素制备糖尿病模型。注射后48 h及5、8 d,断尾采血,测定全血血糖。随机取3次血糖均>16.65 mmol/L的糖尿病大鼠用于移植试验。DMEM稀释每个阳性孔诱导胰岛细胞至100μL,分别移植在12只糖尿病大鼠左侧肾囊内。6只糖尿病模型对照大鼠左侧肾囊内分别注射100 uL DMEM。实验大鼠每10 d断尾测全血血糖1次。在为期67 d的移植试验中,12只诱导胰岛移植大鼠血糖水平从18.93~25.78 mmol/L降至6.32~11.47 mmol/L。除2只死于移植感染外,其余存活了54~67 d。6只糖尿病对照大鼠血糖水平持续在18.73~25.96 mmol/L。1只感染死亡,5只存活了16~42 d。6只正常对照大鼠血糖基本保持在3.56~5.83 mmol/L,死亡率为0。结果表明,人单克隆胰腺干细胞体外定向诱导能分化为包含大量β细胞的功能性胰岛,移植这些诱导胰岛能降低糖尿病大鼠血糖水平,延长其寿命。
Human monoclonal pancreatic stem cells (1.0 × 10 4 cells / well) were seeded in gelatin-coated 6-well plates and induced in vitro to differentiate into islets containing large numbers of β cells. Dithizone staining was positive. RT-PCR detection, transcription of insulin expression. Thirty Sprague-Dawley rats were used as control, and 6 rats were injected intraperitoneally with 2% streptozotocin to prepare diabetic model. At 48 h and 5 and 8 d after injection, the blood was taken off the tail and the whole blood glucose was measured. Three diabetic rats with blood glucose> 16.65 mmol / L were randomly selected for transplantation test. Each positive hole was induced by DMEM to induce islet cells to 100 μL and were transplanted into the left renal capsule of 12 diabetic rats, respectively. Six diabetic rats were injected with 100 uL DMEM into the left renal capsule respectively. Experimental rats were dosed every 10 days to determine the whole blood glucose once. In the 67-day transplantation experiment, the blood glucose level of 12 rats with islet transplantation decreased from 18.93 ~ 25.78 mmol / L to 6.32 ~ 11.47 mmol / L. Except two died of transplanted infection, the rest survived for 54-67 days. Blood glucose levels in 6 diabetic control rats continued at 18.73 ~ 25.96 mmol / L. One died of infection, and five survived for 16-42 days. The blood glucose of 6 normal control rats remained at 3.56 ~ 5.83 mmol / L and the mortality rate was 0. The results showed that human pancreatic stem cells can differentiate into functional islets containing a large number of β cells in vitro. Transplantation of these islets can reduce the blood glucose level and prolong the life span of diabetic rats.