【摘要】：肝干細胞在理論研究和臨床應用都有重要意義。一方面，，肝干細胞可能參與肝臟嚴重損傷后的再生與修復，因此，肝干細胞的增殖與分化及其機理的研究，有助于闡明肝臟的發(fā)育機制。另一方面，肝干細胞具有強大的增殖能力，并能分化為肝細胞，這將為肝病的細胞移植、組織工程和基因治療帶來新的希望。肝組織是否存在肝干細胞的問題一直在爭論，由于肝干細胞缺乏特異性表面標記物，給肝干細胞的分離和鑒定帶來很大困難。成年肝組織的肝干細胞數(shù)量很少，且處于休眠期，要獲取足量且存活率高的肝干細胞，必須建立良好的激活模型。 目的 建立大鼠肝卵圓細胞增殖模型，進一步進行肝干細胞的分離、純化和鑒定。 方法 選擇81只體重200±20g的雄性Wistar大鼠，隨機分為對照組和模型組。采用2-AAF+2／3肝切除方法(2-acetylaminofluorene+two thirds partial hepatectomy，2-AAF+2／3PH)建立肝卵圓細胞增殖模型。使用不同劑量的2—AAF通過胃管灌喂，5天后行肝2／3切除，術后第二天各組按各自劑量繼續(xù)灌喂，并于術后第4h、4d、8d、12d、16d不同的時間點取肝組織，光鏡及電鏡下觀察肝干細胞增殖情況，并行免疫組織化學染色。利用膠原酶原位灌注及Percoll密度梯度離心分離大鼠肝卵圓細胞，并把分離細胞置于37℃5％ CO_2孵箱中培養(yǎng)，細胞在倒置顯微鏡、電鏡下觀察細胞特點，免疫組織化學染色鑒定肝卵
[Abstract]:Liver stem cells are of great significance in both theoretical research and clinical application. On the one hand, liver stem cells may be involved in the regeneration and repair of the liver after severe injury. Therefore, the study of the proliferation and differentiation of hepatic stem cells and its mechanism will help to clarify the mechanism of liver development. On the other hand, liver stem cells have the ability to proliferate and differentiate into hepatocytes, which will bring new hope for cell transplantation, tissue engineering and gene therapy of liver disease. The question of whether liver stem cells exist in liver tissue has always been debated. Due to the lack of specific surface markers of liver stem cells, it is difficult to isolate and identify liver stem cells. The number of liver stem cells in adult liver tissue is very small and it is in dormancy stage. To obtain enough liver stem cells with high survival rate, a good activation model must be established. Objective to establish rat liver oval cell proliferation model and further isolate, purify and identify hepatic stem cells. Methods 81 male Wistar rats weighing 200 鹵20 g were randomly divided into control group and model group. Liver oval cell proliferation model was established by 2-AAF 2 / 3 hepatectomy (2-acetylaminofluorene two thirds partial hepatectomy,2-AAF 2/3PH). After 5 days of intragastric administration of 2-AAF, 2 / 3 hepatectomy was performed in each group, and the liver tissues were taken at different time points at 4 h, 4 d, 8 d, 12 d and 16 d after operation. The proliferation of hepatic stem cells was observed under light microscope and electron microscope, and immunohistochemical staining was used. Rat hepatic oval cells were isolated by in situ perfusion of collagenase and Percoll density gradient centrifugation. The isolated cells were cultured in 5% CO_2 incubator at 37 鈩

本文編號：2211801