人胰岛细胞制备过程中细菌性污染的监测与干预  

Monitoring and intervention on bacterial contamination in preparing human islet cells

作　　者：倪艳霞[1] 蔡德鸿[1] 翁建平[2] 曾龙驿[3] 张桦[1] 龙军[4] 

机构地区：[1]南方医科大学内分泌科,广东省广州市510280 [2]中山大学附属第一医院内分泌科,广东省广州市510080 [3]中山大学附属第三医院内分泌科,广东省广州市510630 [4]南方医科大学检验科,广东省广州市510280

出　　处：《中国临床康复》2006年第29期41-43,共3页Chinese Journal of Clinical Rehabilitation

基　　金：广东省科技厅重大科研基金项目(B30201)~~

摘　　要：目的:在胰岛细胞制备过程中监测并预防性使用敏感抗生素,防止、清除制备过程中出现的污染,保证用于移植的胰岛细胞质量,以期减少移植后感染的发生,确保移植进一步用于临床的效果。方法:体外分离人胰岛细胞实验是于2004-11/2005-10在广州中山大学附属第三医院中心实验室进行。①成人胰岛细胞的分离纯化及其过程中细菌性污染的监测:胰腺取自成人脑死亡尸体,生前无糖尿病及相关病史(家属知情同意),胰腺进行修剪、灌注后,胰岛细胞消化收集中;COBE-2991细胞纯化机纯化胰岛细胞后;体外培养48h后6个步骤取样进行细菌性污染的鉴定,并作药敏实验。②在同样的制备过程中,分别采用青霉素+链霉素+两性霉素B或泰能+两性霉素B进行干预,未施加干预措施作为对照组。③胰岛细胞的功能测定:采用体外胰岛素释放试验,测定纯化培养后未被污染的胰岛细胞功能。结果:①3组不同处理的胰岛细胞(纯化培养后)的污染率:未用抗生素的对照组为100%、青霉素+链霉素+两性霉素B组为50%,泰能+两性霉素B组为0,3组污染率差异有显著性(P<0.05)。②取培养后未污染的胰岛细胞行体外胰岛素释放试验结果:青霉素+链霉素+两性霉素B组、泰能+两性霉素B组两组胰岛细胞高糖时胰岛素释放量分别是低糖的2.41和2.36倍(t=4.76,P<0.05),提示青霉素+链霉素+两性霉素B组、泰能+两性霉素B组胰岛细胞分泌功能良好。两组间的胰岛素敏感指数(ISI)在统计学上差异也无显著性(t=0.31,P=0.76),说明两组抗生素对胰岛细胞功能的影响无差异。结论:胰岛细胞制备过程中细菌性污染严重。泰能+两性霉素B的联合应用能有效清除胰岛细胞制备过程中出现的细菌性污染,经泰能+两性霉素B处理的胰岛细胞行胰岛素释放试验表明分泌功能良好。AIM： To monitor and prophylactically use sensitive antibiotics as well as prevent and eradicate the contamination in the process of islet-cell preparation, so as to ensure the quality of islet cells for transplantation, reduce the incidence of post transplant infections and guarantee the effects of transplantation in clinic. METHODS： The experiment of human islet cell isolation /n vitro was conducted in the Central Labomtory of Third Hospital Affiliated to Sun Yatsen University between November 2004 and October 2005.①The monitoring on bacterial contaminations in the procedures of isolation and purification of adult islet cells： the pancreas was obtained from adult body. Subjects had no diabetes or relevant bistory of disease before died （the relatives all knew and agreed with the items）. The islet cells were digested and collected after trimming and perfusion. Islet cells were purified with COBE-299 cell purification machine and then cultured in vitro for 48 hours. The samples were identified for bacterial contaminations in 6 steps, meanwhile, susceptibility test were performed. ②In the process of same preparation, penicillin ＋ streptomycin ＋ amphotericin B and tienam ＋ ampbotericin B were adopted respectively for interventions. Samples without interventions were taken as the control group. ③Functional examination of islet cells： in vitro insulin releasing test was used to determine the secretory functions of uncontaminated islet cells after purification and culture. RESULTS： ①The contamination rate of 3 groups after treatments with various antibiotics were： it was 100% in the control group （without administration of any antibiotics）, 50% in penicillin ＋ streptomycin ＋ amphotericin B group and 0% in tienam ＋ ampbotericin B group respectively. There was significant difference among 3 gToups（P〈0.05）. ②nsulin releasing test of uncontaminated cultured islet cells/n vitro： The insulin releasing＇ volume of both groups under high glucose condition was 2.41 and 2.36 t

关 键 词：胰岛细胞 细胞培养技术 抗生素类 复合 

分 类 号：R587[医药卫生—内分泌]