【摘要】 目的探讨在不同培养基条件下,不同年龄大鼠的海马脑片在长期培养过程中的活性改变及微管相关蛋白tau表达的动态变化。方法制备1、2、4和8周龄雄性Wistar大鼠海马脑片(400μm),分别使用minimumessentialmedium(MEM)和Dulbecco’smodifiedeaglemedium:nutrientmixture(DMEM/F12)两种不同的培养基进行培养。在培养的21d内以乳酸脱氢酶(LDH)法监测不同培养基条件下海马脑片活性的改变,并在不同时间点选取脑片以免疫印迹法检测tau蛋白含量的改变。结果在海马脑片的长期培养过程中,两种培养基对脑片活性的影响差异无显著性,而培养基DMEM/F12可使脑片中的tau蛋白更持久、更高量地稳定表达,尤其是2和4周龄鼠源海马器官型脑片。结论选取2或4周龄大鼠,应用培养基DMEM/F12更适合于脑片水平tau蛋白的相关研究,在此基础上可望建立阿尔茨海默病理想研究模型。更多还原

【Abstract】 Objective To explore the effect of different culture media on viability and expression of tau protein in organotypic hippocampal slice. Methods Brain slices(400 μm)from 1, 2, 4, and 8 week-old Wistar rats were prepared and cultured in minimum essential medium(MEM)or Dulbecco’s modified eagle medium: nutrient mixture(DMEM/F12)medium respectively for 21 days. Viability of the slices was measured by lactate dehydrogenase(LDH)assay and expression of tau protein was detected by Western blot. Results The viability of the slices was not influenced significantly by the two different culture media, while the expression level of tau protein was significantly higher in DMEM/F12 than in MEM(P < 0.05), especially in the slices from 2 and 4 week-old rats. Conclusion The slices from 2 or 4 week-old rat hippocampi and DMEM/F12 medium may be the preferred choice for tau associated researches. An ideal Alzheimer’s disease model may be established based on the results of these researches.更多还原