CAJ | 학술논문

目的：研究大鼠肝细胞分离的简易方法及长期培养过程中肝细胞形态变化过程。方法：采用体外胶原酶二步灌注法分离大鼠肝细胞，TB染色法计算细胞数及细胞活率。MTT法观测新生牛血清对大鼠肝细胞增殖的影响。在含10％新生牛血清及其他附助因子的Williams＇E条件培养基中原代长期培养，并进行形态学的动态观察。结果：平均每只大鼠可获取2．26×108个肝细胞，平均活力为95．6％。新生牛血清浓度与大鼠肝细胞增殖有明显的量效关系(P＜0．01)。在Williams＇E培养基中可存活5～6周并保持正常形态。结论：本方法分离的肝细胞有较高的获取率和活力，适合体外长期原代培养。
목적：연구대서간세포분리적간역방법급장기배양과정중간세포형태변화과정。방법：채용체외효원매이보관주법분리대서간세포，TB염색법계산세포수급세포활솔。MTT법관측신생우혈청대대서간세포증식적영향。재함10％신생우혈청급기타부조인자적Williams＇E조건배양기중원대장기배양，병진행형태학적동태관찰。결과：평균매지대서가획취2．26×108개간세포，평균활력위95．6％。신생우혈청농도여대서간세포증식유명현적량효관계(P＜0．01)。재Williams＇E배양기중가존활5～6주병보지정상형태。결론：본방법분리적간세포유교고적획취솔화활력，괄합체외장기원대배양。
Objective :To study a simplified method of isolation of rat hepatocytes and to observe the pro-cess of cell morphology in long-term culture. Methods :Rat hepatocytes were isolated by a single two-stepperfusion method. The yield and viability were assessed by trypan blue exclusion. [3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide] (MTT) was used to test the effect of serum concentration of newborn calf serum on the proliferation of hepatocytes. Hepatocytes were inoculated in the culture mediumconsisted of Williams' E supplemented with insulin,dexamethasone and 10％ new born calf serum. Themorphologic change of cultured hepatocytes was observed. Results:The average yield of hepatocytes was 2.26× 108 cells per rat, with an average viability of 95.6％. New born calf serum had strong biological activi-ty to stimulate the proliferation of hepatocytes and there was close-effect relationship followed by the in-crease of new born calf serum concentration. The rat hepatocytes can be cultured for 5～ 6 weeks withpreservation of normal morphologic appearance. Conclusion:The rat hepatocytes isolated by the abovemethod have high yields and viability and can be long-term cultured in vitro.