经心脏逆行置管灌注法在小鼠原代肝细胞分离中的应用

发布时间：2018-06-20 14:31

本文选题：原代肝细胞 + 胶原酶灌注法　；参考：《四川大学学报(医学版)》2014年01期

【摘要】：目的观察采用经心脏逆行置管灌注肝脏的方法分离小鼠原代肝细胞的效果。方法对传统的分离原代肝细胞方法(Seglen两步胶原酶灌注法)进行改良,即从小鼠右心房的裂口处经肝上下腔静脉向下方置管,并以输液器滴注含体积分数10%胎牛血清的胶原酶灌注液进行肝脏灌注。评价此法与传统方法的置管成功率及所得肝原代细胞的数量及活力。结果改良组总的置管操作成功率高达95%,且在一次置管成功率上改良组高于传统组(94.7%vs.68.8%,P0.05)。改良组小鼠肝脏灌注均匀,肝细胞产量高达1.07×106/g小鼠体质量,细胞活力平均为92.16%,两者均高于传统组(分别为0.99×106/g小鼠体质量及86.44%,P0.05或P0.01)。结论经心脏逆行置管灌注法分离小鼠原代肝细胞简便易学,重复性高,且能保证分离所得的肝细胞产量和质量。
[Abstract]:Objective to observe the effect of retrograde perfusion of murine hepatocytes with retrograde catheterization of the heart. Methods Seglen's two-step collagenase perfusion method was modified, that is to say, a tube was inserted from the opening of the right atrium of the mouse through the superior and inferior hepatic vena cava. The liver was infused with collagenase perfusion solution containing 10% fetal bovine serum. To evaluate the success rate of catheterization and the number and activity of primary hepatocytes obtained by this method and the traditional method. Results the total success rate of catheterization in the modified group was 95%, and the improvement group was higher than the traditional group (94.7vs.68.8% P0.05). In the modified group, the liver was perfused evenly, the hepatocyte output was as high as 1.07 脳 106 / g and the average cell activity was 92.16, both of which were higher than those in the traditional group (0.99 脳 106 / g and 86.4444 / g, P0.05 or P0.01, respectively). Conclusion the method of retrograde catheterization through the heart is simple and easy to study and has high reproducibility and can ensure the yield and quality of the isolated hepatocytes.
【作者单位】：四川大学华西医院肝胆胰外科;
【基金】：四川省科技厅科技支撑计划项目(No.2010FZ0016)资助
【分类号】：R318

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