泡球蚴囊液对原代培养大鼠肝细胞增殖影响的初步研究

发布时间：2018-01-11 20:33

本文关键词：泡球蚴囊液对原代培养大鼠肝细胞增殖影响的初步研究　出处：《新疆医科大学》2008年硕士论文　论文类型：学位论文

【摘要】： 目的1.成功建立大鼠肝细胞的无血清原代培养体系。2.探讨泡球蚴囊液对体外无血清原代培养大鼠肝脏细胞增殖的影响。方法1.以Wistar大鼠作肝细胞供者,采用胶原酶肝脏原位灌流消化法分离大鼠肝细胞,并以1×106的密度接种于I型胶原铺底的直径35mm的培养皿,分别采用血清和无血清培养24h和48h,并在肝细胞形态学和肝细胞增殖指数上对两者进行比较,建立大鼠肝细胞无血清原代培养体系。2.大鼠肝细胞无血清培养体系建立后,分别用无菌的泡球蚴囊液及无血清培养液与大鼠肝细胞共培养24h和48h,培养期间用荧光倒置显微镜观察大鼠肝细胞的形态学改变,用流式细胞仪测定大鼠肝细胞的细胞周期;同时将分离所得的大鼠肝细胞以1×105的密度接种于I型胶原铺底的96孔培养板,无血清培养后,采用MTT法检测泡球蚴囊液对大鼠肝脏细胞是否有毒性作用。结果1.大鼠肝细胞无血清培养体系的建立:大鼠肝细胞活细胞分离存活率90%,与血清原代培养的肝细胞相比,无血清原代培养的肝细胞生长良好,24h和48h肝细胞增殖指数略微增加,差异无统计学意义(P0.05)。2.与对照组相比,与泡球蚴囊液共培养的肝细胞贴壁生长良好;MTT法显示泡球蚴囊液对肝细胞无细胞毒性作用;泡球蚴囊液处理24小时后肝细胞增殖指数(PI)由82.0%±6.0%降至49.35%±4.0%;处理48小时后肝细胞增殖指数(PI)由67%±10%降低至27.61%±6%,差异有统计学意义(P0.05)。结论1.成功建立大鼠肝细胞无血清培养体系。2.泡球蚴囊液对于体外培养的大鼠肝细胞无毒害作用,但对肝细胞的增殖有一定的抑制作用。
[Abstract]:Objective 1. To successfully establish the serum-free primary culture system of rat hepatocytes. 2. To investigate the effect of hydatid cyst fluid on the proliferation of rat liver cells in vitro. Methods 1. Wistar rats were used as liver cells. Cell donor. Rat hepatocytes were isolated by in situ perfusion digestion with collagenase and inoculated with a density of 1 脳 10 ~ 6 in a culture dish with a diameter of 35 mm on the bottom of type I collagen. Serum and serum-free culture were used for 24 h and 48 h, respectively, and the morphology and proliferation index of hepatocytes were compared. The primary culture system of rat hepatocytes without serum was established. After the establishment of serum-free culture system, rat hepatocytes were co-cultured with rat hepatocytes in aseptic hydatid cyst fluid and serum-free medium for 24 h and 48 h, respectively. The morphological changes of rat hepatocytes were observed by fluorescence inverted microscope and the cell cycle of rat hepatocytes was measured by flow cytometry. At the same time, the isolated rat hepatocytes were inoculated with 1 脳 105 density on the 96-well culture plate covered with collagen type I, and cultured without serum. MTT method was used to detect the toxic effect of hydatid cyst fluid on rat liver cells. Results 1. Establishment of serum-free culture system of rat hepatocytes: survival rate of rat hepatocytes was 90%. Compared with primary cultured hepatocytes without serum, the proliferation index of hepatocytes in serum-free primary culture increased slightly after 24h and 48h. Compared with the control group, the hepatocytes co-cultured with hydatid cyst fluid had a good adherent growth. MTT assay showed that hydatid cyst fluid had no cytotoxic effect on hepatocytes. The proliferative index (Pi) of hepatocytes decreased from 82.0% 卤6.0% to 49.35% 卤4.0 after 24 hours treatment with hydatid cyst fluid. After 48 hours of treatment, the proliferation index of hepatocytes decreased from 67% 卤10% to 27.61% 卤6%. The difference was statistically significant (P 0.05). Conclusion 1. Successful establishment of serum-free culture system of rat hepatocytes. 2. Hydatid cyst fluid has no toxic effect on rat hepatocytes cultured in vitro. 2. But it can inhibit the proliferation of hepatocytes.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R363

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