乳鼠窦房结细胞取材与纯化培养方法的探讨

Purification and Primary Culture of the Sinoatrial Node Cells Isolated From the Neonatal Rat.

本研究旨在建立一套可靠的窦房结细胞取材分离、纯化培养技术。取Wistar乳鼠的窦房结组织进行原代细胞培养 ,随机分两组 :常规方法组与差速贴壁结合 5 溴脱氧尿核苷 (BrdU)处理组。结果 :①在培养的窦房结细胞中观察到三种不同形态的细胞 ,即梭形细胞、三角形细胞与不规则形细胞。其中梭形细胞最多 ,且形态结构与电生理特征符合窦房结起搏细胞的特点。三角形细胞与心房肌三角形细胞特征相似。②常规方法培养中窦房结梭形细胞所占比例为 45 % ,而差速贴壁技术结合BrdU处理的纯化培养中梭形细胞达 6 8%左右 (P <0 .0 1)。结论 :用差速贴壁结合BrdU处理的纯化培养可明显提高窦房结梭形细胞的比例 。

This study was designed to develop a reliable method of primary culture of the isolated sinoatrial node (SAN) cells.The isolated neonatal rat SAN cells were randomly devided into two groups:routine culture group and differential attachment and 5′ Bromo 2′ deoxyuridine BrdU treated group.Among the cultured cells of SAN,we observed three different configuration cells:spindle cells,triangle cells and irregular cells.The greatest proportion of cells were spindle cells.The greated proportion of cells were spindle cells whose configuration,structure and electrophysiological characteristics accorded with that of pacemaker cell of SAN.However,triangle cells isolated from SAN were similar with triangle cells of atrial muscle and the second proportion were triangle cell.The proportion of spindle cells increased from 45% among the cells cultured with routine method to 68% among cultured with differential attachment and BrdU treated method.Conclusions:The culture method of differential attachment and BrdU treated can significantly increase the proportion of spindle cells and is a reliable method of primary culture of the isolated SAN cells.[Chinese Journal of Cardiac Pacing and Electrophysiology,2000,14(3):188～190]