采用改良的胰腺三管插管法分离成人胰岛的实验研究  

Improved islet isolation by three-cannula method for collagenase infusion

作　　者：程颖[1] 石蕊[1] 李桂臣[1] 吴刚[1] 刘树荣[1] 孟一曼[1] 赵宁[1] 刘永锋[1] 

机构地区：[1]中国医科大学附属第一医院普通外科教研室器官移植科,沈阳,l10001

出　　处：《中华器官移植杂志》2011年第10期630-632,共3页Chinese Journal of Organ Transplantation

基　　金：基金项目：卫生部行业专项基金（2（）0802003）;国家科技支撑课题（2（）08BAl60806）;辽宁省博士启动基金（20081051）

摘　　要：目的探讨胰腺插管方式对成人胰岛分离及纯化的影响。方法共对17例成人胰腺进行了胰岛的分离和纯化。采用改进的腹部器官联合快速切取技术获取胰腺，分别采用标准法（3例）、单管法（11例）和三管法（3例）对胰腺进行灌注。标准法是将胰腺从胰颈处完全切断，沿主胰管分别向胰头和胰尾插管，主胰管人十二指肠处予以结扎。单管法为采用加长插管自主胰管插入，直至胰尾。三管法是在胰颈背侧切开胰腺至主胰管，经主胰管分别向胰头和胰尾方向插管，在主胰管进入十二指肠处插第3根插管。采用胶原酶LibarseHI消化，Ficoll连续密度梯度离心法纯化。双硫腙染色，鉴定胰岛的纯度，并计算胰岛当量（IEQ）。丫啶橙／溴乙啶荧光染色，计数活细胞百分率。体外葡萄糖刺激试验鉴定胰岛功能。结果标准法的灌注量平均为0．71ml／g胰腺，单管法的灌注量平均为0．96ml／g胰腺，三管法的灌注量平均为1．24ml／g胰腺，明显多于前两种方法（P〈0．05）。标准法的胰岛收获量平均为1914IEQ／g胰腺，单管法为2270IEQ／g胰腺，三管法为2514IEO．／g胰腺，单管法和三管法明显高于标准法（P〈0．05）；其胰岛纯度／活性分别为74％／79．3％、75．6％／79．4％和78．3％／84．0％，三者问的差异无统计学意义。标准法所获得的胰岛胰岛素释放指数平均为3．46，单管法为4．74，三管法为5．27，单管法和三管法明显高于标准法（P〈0．05）。结论不同的插管灌注方式对成人的胰岛分离有一定影响，三管法有利于提高胰腺灌注量，增加胰岛的收获量。Objective Pancreas perfusion is an essential step in human islet isolation. To develop the new methods for introductal canulation, collagenase infusion and to observe their effects on islets isolation. Methods A total of 17 pancreases were digested from March 2005 to April 2010. The pancreases were distended by three methods： the standard method （n = 3）, the onecannula method （n = 11 ） and three-cannula method （n = 3）. In the standard group, the pancreases were completely cut into half at the mid-body. Two catheters were inserted into the main duct： one directed toward the tail and the other to the head. In the one-cannula method group, a long tube was inserted into the duct at the head, advancing to the tail. In the three-cannula method group, pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct： one at the head （the first catheter） and two at the mid-body, one toward the tail （the second catheter） and the other toward the head （the third catheter）. The pancreases were digested by improved Ricordi technique. Ficoll continuous density/grads centrifuge method was performed to purify the islets. DTZ staining was adopted to identify jslets and count islet equivalent （IEQ）. AO/EB fluorescence examination was used to count active islet percentage. Static glucose stimulating test （SGS） in vitro was designed to estimate islet function and calculate SI. Results The distension volume of the three- cannula method group was 1.24 ml/g pancreas, and higher than the other groups （for the standard group： 0. 71 ml/g pancreas;for one-cannula method group.. 0. 96 ml/g pancreas, P〈0. 05）. The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g, which was significantly more than that in the standard group （1914 IEQ/g pancreas, P〈0. 05）. The purity and viability of the islets were 74 %/79. 3 %, 75.6 %/79. 4 % and 78. 3 %/ 84. 0 % respectively in the three

关 键 词：胰腺 插管法 分离 胰岛 

分 类 号：R657.5[医药卫生—外科学]