【摘要】：目的 通過臍血干細胞移植在體內(nèi)誘導分化為肝細胞的實驗研究,進一步為臍血干細胞向肝細胞的分化提供證據(jù),證實臍血干細胞的可塑性,為將臍血干細胞移植應用于治療終末期肝病的研究奠定基礎。 方法 ① 使用采血袋法采集臍血標本,分別采用改良HES(羥乙基淀粉)沉降分離法和Ficoll分離法兩種方法分離臍血干細胞,比較分離前后有核細胞(nuclear cell,NC)數(shù)、分離后CD34~+細胞數(shù)、分離后有核細胞回收率及臺盼藍拒染率,了解分離后有核細胞的數(shù)量和質(zhì)量,為臍血移植創(chuàng)造條件。 ② 選用BALB/C裸鼠,將其隨機分成實驗組和對照組,實驗組再根據(jù)輸注有核細胞數(shù)的不同分成A、B、C三組。采用Co~(60)治療儀γ-射線對實驗組和對照組進行亞致死劑量350cGy照射,照射后3小時內(nèi)在無菌條件下由裸鼠尾靜脈輸入分離的人新鮮臍血有核細胞。實驗A、B、C三組分別輸入NC1.0×10~7個/只、2.0×10~7個/只、3.0×10~7個/只,對照組經(jīng)尾靜脈注入等體積無菌生理鹽水。移植后第1、2、3、4和8周分別檢測實驗組和對照組外周血白細胞,移植后第4、6和8周通過流式細胞儀檢測外周血人源性CD34~+和CD45~+細胞水平,了解人鼠細胞嵌合情況。移植后一個月對實驗組A、B和對照組裸鼠分別按0.4ml/kg注射用玉米油稀釋到100μl的四氯化碳(carbon tetrachloride,CCl_4),實驗組C注射等體積生理鹽水。通過CCl_4造成肝臟損傷,誘導臍血干細胞歸巢到
[Abstract]:Objective to provide evidence for the differentiation of umbilical cord blood stem cells into hepatocytes by transplantation of umbilical cord blood stem cells in vivo, and to confirm the plasticity of cord blood stem cells. To lay a foundation for the application of umbilical cord blood stem cell transplantation in the treatment of end-stage liver disease. Methods (1) Cord blood samples were collected by blood bag method. Two methods, modified HES (hydroxyethyl starch) sedimentation method and Ficoll method, were used to isolate umbilical cord blood stem cells. The number of nucleated cells (nuclear cell,NC) before and after isolation was compared. The number of CD34~ cells after separation, the recovery rate of nucleated cells and trypan blue rejection rate after separation, the quantity and quality of nucleated cells after separation were understood to create conditions for cord blood transplantation. 2 BALB/C nude mice were randomly divided into experimental group and control group. The experimental group was divided into three groups according to the number of nucleated cells. The experimental group and control group were irradiated with sublethal dose of 350cGy with Co~ _ (60) therapeutic instrument. Within 3 hours after irradiation, human fresh cord blood nucleated cells were injected into nude mice 'tail vein in aseptic condition. NC1.0 脳 10 ~ 7 / mouse, 2.0 脳 10 ~ 7 / mouse and 3.0 脳 10 ~ 7 / mouse were injected into the control group respectively. The control group was injected with the same volume of aseptic saline through the caudal vein. Peripheral white blood cells (PBWs) in the experimental group and the control group were detected at 4 and 8 weeks after transplantation. The levels of human derived CD34~ and CD45~ cells were detected by flow cytometry at the 6th and 8th weeks after transplantation. The chimerism of human mouse cells was investigated. One month after transplantation, the experimental group and the control group were diluted to 100 渭 l carbon tetrachloride (carbon tetrachloride,CCl_4) with corn oil for 0.4ml/kg injection, while the experimental group C was injected with the same volume of normal saline. Liver injury induced by CCl_4 induces umbilical cord blood stem cells to homing
【學位授予單位】：四川大學
【學位級別】：博士
【學位授予年份】：2005
【分類號】：R329

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