原代培养乳鼠窦房结细胞的形态和kir2.1蛋白表达

目的 研究乳鼠窦房结细胞中kir2.1蛋白表达水平,建立一套可靠的乳鼠窦房结定位、细胞培养与鉴定技术,为进一步心脏生物起搏实验奠定基础.方法 在石蜡切片中用HE染色、嗜银染色、髓鞘染色和Mallory磷钨酸-苏木素染色法(PTAH)进行乳鼠窦房结定位和形态学观察;取SD乳鼠的窦房结组织进行原代细胞培养,观察培养细胞的形态和搏动频率,免疫组织化学检测其kir2.1蛋白表达.结果 综合嗜银染色、髓鞘染色和PTAH染色3种方法可准确定位乳鼠窦房结组织.在培养的窦房结细胞中观察到梭形细胞、三角形细胞和不规则形细胞3种有自发性收缩的细胞,其中梭形细胞最多且形态结构特点和搏动频率符合起搏细胞的特点,三角形和不规则细胞与心房肌细胞特征相似.乳鼠窦房结细胞的kir2.1蛋白表达低于心房和心室肌细胞.结论 综合嗜银染色、髓鞘染色和PTAH染色3种方法可以准确定位SD乳鼠窦房结,乳鼠窦房结细胞中kir2.1蛋白表达水平低于心房和心室肌细胞.

Primary cultured neonate rat sinus node cells: morphology and expression of kir2.1 protein

Objective To investigate the expression of kir2.1 protein in primary cultured sinus node cells and establish a reliable technique to locate,culture and characterize neonatal rat sinus node cells.Methods In paraffin sections,the location and morphology of the neonatal rat sinus node cells were observed by HE staining,silver nitrate staining,myelin staining and phosphotungstic acid-hematoxylin(PTAH) staining.Primary cell culture from the neonatal rat sinus node was conducted to observe the spontaneous contraction frequency,cell morphology and kir2.1 protein expression.Results Combination of the 3 staining methods allowed accurate localization of the sino-atrial nodal(SAN) tissue,and among the cultured cells in the SAN,at least 3 distinct types of cells with spontaneous contraction were observed.The majority of the contracting cells were spindle cells and their construction and impulse frequency indicated their identity as pacemaker cells,while the triangular and irregular cells resemblesd the atrial musclle cells.A lower expression level of kir2.1 protein was detected in SAN cells than in the atrial and ventricular myocytes of the neonatal rats.Conclusion Combination of silver nitrate staining,myelin staining and PTAH staining identifies the exact location of the sinus node tissue,and cultured sinus node cells have lower expression of kir2.1 protein than the atrial and ventricular myocytes of neonatal rats.