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100 ml UW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0.5～4 h.纯化前收集的胰岛数量平均为(376.7±50.2)×103 IEQ,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.
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 solution. First, liberase was perfused through the main pancreatic duct to distend the whole pancrease. Then the pancrease was cut into 5-8 segments and put into the Ricordi chamber, and shaken and digested under 37℃. The collected islets were dissolved in 100 ml UW solution for 30 rain. The Ficoll solution was pumped into COBE2991, then UW/islet solution and 50 ml wash solution （M199）. After purification, the islet number and purity was calculated, and the activity of the islets was assayed. Results Hot ischemia time was 0-1 rain, and cold ischemia time was 0. 5-4 h. The mean islet number collected before purification was （376. 7 ± 50. 2） × 10^3 IEQ, and after purification it was （378.6 ± 56. 7） × 10^3 IEQ. The mean islet 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. The 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 for clincal transplantation.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2005年第8期467-469,共3页
Chinese Journal of Organ Transplantation
基金
2003年黑龙江卫生厅国际科技合作项目(WC03308)
