大鼠海马神经元培养与鉴定

探索适用于膜片钳记录的大鼠海马神经元的分离及原代培养方法，并检测其电生理特性。采用钝性分离1日龄SD大鼠双侧海马，经酶消化及吸管吹打，用含胎牛血清、马血清的DMEM体外培养，并经阿糖胞苷处理24h，第9—15d用于膜片钳全细胞模式检测神经元电生理特性。结果显示培养的海马锥体神经元表面光洁，膜片钳记录封接成功率90％，具有正常神经元的电生理及反应特性。表明本分离及原代培养方法，有益于大鼠海马锥体神经元生长，并适用于膜片钳记录。

Cultures and identification of hippocampal pyramidal neurons in rats

s: To develop the methods of isolating and cultivating of hippocampal pyramidal neurons of Sprague-Dawley rats and to detect electrophysiological properties of the neurons. Primary cultured hippocampal cells were carefully prepared from the neurons SD rats of one day age, After chopped into 0 4-mm cubes, the cubes were mechanically dispersed with pipettes in the presence of trypsin (0 25%,g/L) and DNase (0 004%,g/L). Hippocampal cells were cultured in DMEM enriched with 10% fetal bovine serum and 5% horse serum. Three days later, cytosine arabinoside ( 5 mol/L) were added into the culture medium for another 24 hours. The electrophysiological properties of pyramidal neurons in hippocampus were examined in whole-cell mode in the period from day 9 to day 15. The sealing rate was over 90%. Normal electrophysiological properties and reaction of those cells were shown in the test. The methods of isolation and primary cultivation of hippocampal pyramidal neurons of SD rats improved the growth of neurons and so might be used in patch-clamp recording.