【摘要】 目的通过机械分离胰腺的方法,提高成人胰岛数量和质量。方法切取8个成人尸体胰腺,并保存于4℃UW液中。首先经主胰管插管灌注复合胶原酶溶液,使胰腺膨胀,然后将胰腺剪切成5~8块,放入Ricordi室中,于37℃下机械震荡消化。收集的胰岛溶于100mlUW液中,放置30min。先将Ficoll液泵入COBE2991细胞分离仪中,然后泵入UW/胰岛制剂,最后加入M199液冲洗。收集纯化的胰岛,计算胰岛当量(IEQ)和纯度,检测胰岛的活性。结果热缺血时间为0~1min,冷缺血时间0.5~4h。纯化前收集的胰岛数量平均为(376.7±50.2)×103IEQ,纯化后为(378.6±56.7)×103IEQ;胰岛收获率平均为(86.8±9.4)%,镶嵌的胰岛平均占(11.0±2.7)%;收获的胰岛纯度平均为(88.3±5.1)%;经AO-PI染色后检测胰岛的活度为(91.3±7.8)%。胰岛素分泌指数平均为5.87±0.81。结论通过机械分离胰腺的方法,可以获得更多的胰岛数量和更高的胰岛活性,适合用于临床移植。更多还原

【Abstract】 Objective To improve adult islet isolation quality with mechanization method. Methods Pancreases were abtained from 8 adult human cadaver donors and put into 4℃ UW s olution. First, liberase was perfused through the main pancreatic duct to disten d the whole pancrease. Then the pancrease was cut into 5～8 segments and put int o the Ricordi chamber, and shaken and digested under 37℃. The collected islets were dissolved in 100 ml UW solution for 30 min. The Ficoll solution was pumped into COBE2991, then UW/islet solution and 50 ml wash solution (M199). After puri fication, the islet number and purity was calculated, and the activity of the is lets was assayed. Results Hot ischemia time was 0～1 min, and cold ischemia time was 0.5 ～4 h. The mea n islet number collected before purification was ( 376.7 ± 50.2 )×103I EQ, and after purification it was ( 378.6 ± 56.7 )×103IEQ. The mean is let recovery rate was ( 86.8 ± 9.4 )% and recovery purity was ( 88.3 ± 5.1 )% respectively. The mean trapped islets reached ( 11.0 ± 2.7 ) %. The islet activity reached ( 91.3 ± 7.8 )% through AO-PI staining. T he mean insulin secretory index was 5.87 ± 0.81 . Conclusions We could acquire more purified islets having better activity. Through the Edmonton protocol the isolated isets were enough fo r clincal transplantation.更多还原