【摘要】 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。更多还原

【Abstract】 The monoclonal human pancreatic stem cells(about 1.0×104 cells/well) were cultured in 6-well plates.After proliferated for 3 days,these cells were induced to differentiate into pancreatic islets-like that included a lot of β cells.The induced functional pancreatic islets were positive with dithizone staining,expressed the transcription factor of the insulin by RT-PCR.Total thirty SpragueDawley adult rats were used for transplantation.Six rats as the negative control were individually injected with buffer.Twenty four rats were injected with 2% strptozotocin.On the 48 h,5 th and 8 th days after being injected with the STZ,blood samples were collected by cutting tail,and plasma glucose levels were measured.Rats with plasma glucose levels over 16.65 mmol/L by three time measurements were used as diabetes models for transplantation.The induced pancreatic islets invevery well which showed positive were suspended in 100 μL DMEM,and transplanted into the subcapsular region of the left kidney of twelve diabetes rats.At the same time,the subcapsular region of the left kidney of the other six diabetes model control rats was injected with 100 μL DMEM.The blood glucose levels of the experimental rats were measured in samples collected by cutting tail every 10 days.During 67 day experiment,the blood glucose levels of the twelve rats transplanted with induced pancreatic islets gradually decreased from 18.93-25.78 mmol/L to 6.32-11.47 mmol/L.These rats lived for 54-76 days,except two died 3 days by transplantation infection.Among six diabetes model control rats,five lived for 16-42 days with the blood glucose levels of 18.73-25.96 mmol/L and one died 3 days by transplantation infection.All six normal control rats were alive with the blood glucose levels of 3.54-5.50 mmol/L.These results indicated that the monoclonal human pan creaticstem cells could be in vitro induced into functional islet-like clusters included many β cells;The transplantation of induced pancreatic islets could decrease blood glucose levels of diabetes rats and prolong the life time.更多还原