乳鼠窦房结细胞的原代培养与鉴定

Culture and identification of primary sinoatrial node pacement cells in rat

目的 :建立一套可靠的窦房结细胞培养与鉴定技术。方法 :取Wistar乳鼠的窦房结组织 ,用差速贴壁技术及BrdU处理法进行原代细胞培养 ;对培养细胞进行形态学观察 ,用电生理技术记录细胞的动作电位。结果 :在培养的窦房结细胞中观察到有 3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。结论 :①用差速贴壁技术及BrdU处理法进行乳鼠的窦房结细胞培养 ,是一种可靠的窦房结细胞培养技术。

Objective:To develop a reliable method for culture and identification of primary sinoatrial node (SAN) cells. Methods: The neonatal Wistar rat SAN cells were cultured and purified with the method of differential attachment and BrdU treated. The cultured cells were identified by morphology, spontaneous contraction frequency and ultrastructure observed with Transmission Electromicroscope. The SAN cells were further confirmed by action potential measured with electrophysiological technique. Results: Three distinctly different types of cells were observed among the cultured cells of SAN, which were spindle cells, triangle cells and irregular cells. The spindle cells were the greatest proportion of the cultured cells of SAN. The configuration, structure and electrophysiological character of the spindle cells were accorded with the character of pacemaker cell of SAN. There was no significant difference between triangle cells isolated from SAN and from atrial muscle.Conclusion: The culture method of differential attachment and BrdU treated is a reliable method of culture of the SAN cells. Among the cultured cells of SAN, the spindle cells were pacemaker cells of SAN in rats.