低温微重力培养对大鼠胰岛细胞质量影响的实验
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邢军

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Effect of transplantation of rat islets improved by low temperature microgravity culture
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    摘要:

    目的:研究低温微重力对大鼠胰岛移植质量的影响,以期减少体外培养对胰岛活性和数量的影响,提高胰岛移植质量。方法:将分离纯化的大鼠胰岛分为3组:大鼠新鲜胰岛移植组(于移植前在普通培养基中培养21 d,对照组);实验1组(大鼠新鲜胰岛在37 ℃ RCCS中培养21 d);实验2组(新鲜大鼠胰岛在26 ℃ RCCS中培养14 d后复温至37 ℃继续在37 ℃ RCCS中培养7 d)。 观察各种培养液中的胰岛素含量。并将上述3个实验组不同条件培养的胰岛分别以2000IEQ胰岛移植量植入糖尿病大鼠体内,并观察10周。结果:实验2组的胰岛存活率、胰岛素分泌水平及胰岛素刺激指数均高于对照组和实验1组,差异有统计学意义(P<0.05)。21 d时实验2组胰岛存活率高达(67.4±4.6)%,而对照组和实验1组胰岛存活率分别降至(28.1±3.3)%和(50.3±3.5)%,3组间差异有统计学意义(P<0.05)。给糖尿病大鼠移植实验1,2组的胰岛后均能控制血糖至正常水平,但实验2组胰岛移植对糖尿病大鼠7 d内血糖控制优于实验1组;对照组血糖控制差,3组间两两比较均有统计学差异(均P>0.05)。结论:大鼠胰岛经低温微重力培养后移植,可以明显提高1型糖尿病大鼠的治疗效果。

    Abstract:

    Objective:To observe the effect of transplantation of rat islets which cultured in hypothermia microgravity rotary cell culture system(HRCCS) on rats with diabetes mellitus.Methods :The isolated and purified rat islets were divided into three groups: Temperature (37 ℃)culture for 21 days in flaskculture (control group); temperature(37 ℃)culture for 21 days in RCCS (experimental group1 E1); after low temperature (26 ℃)culture for 14 days in HRCCS, then temperature(37 ℃)culture for 7 days in RCCS (experimental grop 2, E2). Survival rate of islets in three transplanted groups was measured by AOPI doublestaining. Cultured islets were identified by dithizone (DTZ) staining, and insulin contents of different culture liquids were measured by radioimmunoassay. Then the cultured islets was implanted into rat recipients and the transplantation effect was observed for 10 weeks.Results:Survival rate of islants in group E2 (67.4±4.6%) was higher than that in group E1 (50.3± 3.5)% and control group (28.1±3.3)% (all P<0.05). The insulin content and insulin stimulation index in group E2 were higher than those in and group E1 and control group. The survival rate of islets in group E2 (67.4±7.6)% was higher than that in group E1 (28.1±3.3)% and control group (50.3±3.5)% (all P<0.05). Transplantation of group E1 and E2 islets into diabetes rats coulde restored blood glucose to normal in all recipients within one week, and all recipients maintained normal glucosetolerant curve throughout the observation time. But the effect of group E2 islets transptation on didabetes rats was better than that in group E1 islets transplation; and the effect of control group islets transplantion for controlling blood glucose is not so good. Conclusions:Rat islets cultured in 26℃microgravity rotary cell culture system(26℃RCCS) could improve the effect of islet transplantation.

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邢军, 陈艳波, 吴舰宇, 周毅, 宋纯, 宋春芳.低温微重力培养对大鼠胰岛细胞质量影响的实验[J].中国普通外科杂志,2008,17(11):15-110.
DOI:10.7659/j. issn.1005-6947.2008.11.015

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  • 在线发布日期: 2008-11-25