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

  更多相關(guān)文章： 泡球蚴囊液 大鼠肝細(xì)胞 MTT 細(xì)胞增殖

【摘要】： 目的1.成功建立大鼠肝細(xì)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(xì)胞增殖的影響。方法1.以Wistar大鼠作肝細(xì)胞供者,采用膠原酶肝臟原位灌流消化法分離大鼠肝細(xì)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(xì)胞形態(tài)學(xué)和肝細(xì)胞增殖指數(shù)上對兩者進(jìn)行比較,建立大鼠肝細(xì)胞無血清原代培養(yǎng)體系。2.大鼠肝細(xì)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(xì)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(xì)胞的形態(tài)學(xué)改變,用流式細(xì)胞儀測定大鼠肝細(xì)胞的細(xì)胞周期;同時(shí)將分離所得的大鼠肝細(xì)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(xì)胞是否有毒性作用。結(jié)果1.大鼠肝細(xì)胞無血清培養(yǎng)體系的建立:大鼠肝細(xì)胞活細(xì)胞分離存活率90%,與血清原代培養(yǎng)的肝細(xì)胞相比,無血清原代培養(yǎng)的肝細(xì)胞生長良好,24h和48h肝細(xì)胞增殖指數(shù)略微增加,差異無統(tǒng)計(jì)學(xué)意義(P0.05)。2.與對照組相比,與泡球蚴囊液共培養(yǎng)的肝細(xì)胞貼壁生長良好;MTT法顯示泡球蚴囊液對肝細(xì)胞無細(xì)胞毒性作用;泡球蚴囊液處理24小時(shí)后肝細(xì)胞增殖指數(shù)(PI)由82.0%±6.0%降至49.35%±4.0%;處理48小時(shí)后肝細(xì)胞增殖指數(shù)(PI)由67%±10%降低至27.61%±6%,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論1.成功建立大鼠肝細(xì)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(xì)胞無毒害作用,但對肝細(xì)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(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R363

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;中毒及化學(xué)性損害[J];國外科技資料目錄(醫(yī)藥衛(wèi)生);1997年10期

2 唐春;別平;李昆;張玉君;馬正偉;;重組人肝再生增強(qiáng)因子對梗阻性黃疸大鼠肝細(xì)胞線粒體功能的保護(hù)作用[J];消化外科;2006年05期

3 范文今;牛璐;楊鴻武;袁雪芬;解學(xué)魁;;梔芍益肝膠囊對大鼠酒精性肝損傷保護(hù)作用的研究[J];中國比較醫(yī)學(xué)雜志;2007年08期

4 吳其愷;張玲;劉麗嬋;江遠(yuǎn);紀(jì)奕玲;;脂炎消煎劑對大鼠非酒精性脂肪肝肝組織學(xué)的影響[J];河南中醫(yī);2007年11期

5 閔峰;王素美;;丙型肝炎病毒感染離體培養(yǎng)樹，

本文編號(hào)：1411127