精密肝切片技术方法学的建立

目的 :建立一种新的肝脏体外研究手段———精密肝切片技术 ,摸索最佳培养条件。方法 :利用国产振荡切片机 ,制备肝切片 ,建立培养系统 ,以乳酸脱氢酶外漏、谷胱甘肽含量、噻唑蓝还原能力、蛋白含量为指标 ,观察切片厚度、培养液pH值及培养时间对肝切片活力的影响 ;观察抗氧化剂谷胱甘肽 (GSH)、阿魏酸钠 (SF)对CCl4 损伤肝切片活力的影响。结果 :切片厚度 30 0 μm ,培养液pH7.0时 ,肝切片在 4h内活力维持良好。GSH可恢复CCl4 所致肝切片多项指标的变化 ,SF也有一定逆转作用。结论 :精密肝切片技术可用于肝体外研究。切片厚度 30 0 μm ,培养液pH7.0为最适切片与培养条件

Establishment of Technique of Precision-Cut Liver Slice

Objective: To establish the technique of precision cut liver slice(PCLS) and to grope the optimal condition of culturing.Methods: The domestic shaking slicer was used to prepare liver slices. LDH leakage, GSH level, protein content, and MTT reduction were used to assess the slice viability under different pH, thickness of slices and culturing time. And the change of slice viability cultured in the medium with CCl 4 and antioxidants GSH and SF was observed.Results: When the thickness of slices was 300 μm and pH of medium was 7.0, the viability of slices could maintain on a steady level during the first four hours of culture. After treated with CCl 4, the change of slices viability could be inhibited by GSH and SF. Conclusions: The technique of PCLS can be used in study of liver in vitro. Thickness 300 μm and pH 7.0 are the optimal slicing and culturing condition.