CAJ | 학술논문

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0.71 ml/g胰腺,单管法的灌注量平均为0.96 ml/g胰腺,三管法的灌注量平均为1.24 ml/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1914 IEQ/g胰腺,单管法为2270 IEQ/g胰腺,三管法为2514 IEQ/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％,三者间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.
목적 탐토이선삽관방식대성인이도분리급순화적영향.방법 공대17례성인이선진행료이도적분리화순화.채용개진적복부기관연합쾌속절취기술획취이선,분별채용표준법(3례)、단관법(11례)화삼관법(3례)대이선진행관주.표준법시장이선종이경처완전절단,연주이관분별향이두화이미삽관,주이관인십이지장처여이결찰.단관법위채용가장삽관자주이관삽입,직지이미.삼관법시재이경배측절개이선지주이관,경주이관분별향이두화이미방향삽관,재주이관진입십이지장처삽제3근삽관.채용효원매LibarseHI소화,Ficoll련속밀도제도리심법순화.쌍류종염색,감정이도적순도,병계산이도당량(IEQ).아정등/추을정형광염색,계수활세포백분솔.체외포도당자격시험감정이도공능.결과 표준법적관주량평균위0.71 ml/g이선,단관법적관주량평균위0.96 ml/g이선,삼관법적관주량평균위1.24 ml/g이선,명현다우전량충방법(P＜0.05).표준법적이도수획량평균위1914 IEQ/g이선,단관법위2270 IEQ/g이선,삼관법위2514 IEQ/g이선,단관법화삼관법명현고우표준법(P＜0.05)；기이도순도/활성분별위74 ％/79.3％、75.6 ％/79.4％화78.3 ％/84.0％,삼자간적차이무통계학의의.표준법소획득적이도이도소석방지수평균위3.46,단관법위4.74,삼관법위5.27,단관법화삼관법명현고우표준법(P＜0.05).결론불동적삽관관주방식대성인적이도분리유일정영향,삼관법유리우제고이선관주량,증가이도적수획량.
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 one-cannula 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 islets 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 threecannula method group was 1.24 rnl/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 groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).Conclusion The three-cannula method improved collagenase infusion and the islet yields.