本文關鍵詞：泡球蚴囊液對原代培養(yǎng)大鼠肝細胞增殖影響的初步研究　出處：《新疆醫(yī)科大學》2008年碩士論文　論文類型：學位論文

  更多相關文章： 泡球蚴囊液 大鼠肝細胞 MTT 細胞增殖

【摘要】： 目的1.成功建立大鼠肝細胞的無血清原代培養(yǎng)體系。2.探討泡球蚴囊液對體外無血清原代培養(yǎng)大鼠肝臟細胞增殖的影響。方法1.以Wistar大鼠作肝細胞供者,采用膠原酶肝臟原位灌流消化法分離大鼠肝細胞,并以1×106的密度接種于I型膠原鋪底的直徑35mm的培養(yǎng)皿,分別采用血清和無血清培養(yǎng)24h和48h,并在肝細胞形態(tài)學和肝細胞增殖指數(shù)上對兩者進行比較,建立大鼠肝細胞無血清原代培養(yǎng)體系。2.大鼠肝細胞無血清培養(yǎng)體系建立后,分別用無菌的泡球蚴囊液及無血清培養(yǎng)液與大鼠肝細胞共培養(yǎng)24h和48h,培養(yǎng)期間用熒光倒置顯微鏡觀察大鼠肝細胞的形態(tài)學改變,用流式細胞儀測定大鼠肝細胞的細胞周期;同時將分離所得的大鼠肝細胞以1×105的密度接種于I型膠原鋪底的96孔培養(yǎng)板,無血清培養(yǎng)后,采用MTT法檢測泡球蚴囊液對大鼠肝臟細胞是否有毒性作用。結(jié)果1.大鼠肝細胞無血清培養(yǎng)體系的建立:大鼠肝細胞活細胞分離存活率90%,與血清原代培養(yǎng)的肝細胞相比,無血清原代培養(yǎng)的肝細胞生長良好,24h和48h肝細胞增殖指數(shù)略微增加,差異無統(tǒng)計學意義(P0.05)。2.與對照組相比,與泡球蚴囊液共培養(yǎng)的肝細胞貼壁生長良好;MTT法顯示泡球蚴囊液對肝細胞無細胞毒性作用;泡球蚴囊液處理24小時后肝細胞增殖指數(shù)(PI)由82.0%±6.0%降至49.35%±4.0%;處理48小時后肝細胞增殖指數(shù)(PI)由67%±10%降低至27.61%±6%,差異有統(tǒng)計學意義(P0.05)。結(jié)論1.成功建立大鼠肝細胞無血清培養(yǎng)體系。2.泡球蚴囊液對于體外培養(yǎng)的大鼠肝細胞無毒害作用,但對肝細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% 鹵6.0% to 49.35% 鹵4.0 after 24 hours treatment with hydatid cyst fluid. After 48 hours of treatment, the proliferation index of hepatocytes decreased from 67% 鹵10% to 27.61% 鹵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.
【學位授予單位】：新疆醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2008
【分類號】：R363

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