Wang Yanli, Liu Ruxiu, Liu Yu, Li Yang
Department of Cardiology, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2013 Mar;33(3):397-400.
To compare the effects of enzyme digestion with differential attachment and enzyme digestion with differential attachment plus 5'-bromodeoxyuridine (5-BrdU) (conventional) method for isolating sino-atrial node cells from newborn rats for patch clamp studies.
The sino-atrial node tissue were removed from neonatal Wistar rats, digested and separated into single sino-atrial node cells with the above two methods. The obtained cells were observed for morphology, structure, beat frequency, degree of difficulty for patch clamp experiment and the success rate. The atrial cells isolated with enzyme digestion with differential attachment served as the control cells.
The spindle-shaped cells constituted the greatest proportion of the cultured sino-atrial cells, and the cultured atrial cells showed mostly a triangular morphology. Both of the sino-atrial cells separated with the two methods contained mostly spindle-shaped cells, in which spontaneous action potential was recorded. The spindle-shaped cells isolated with enzyme digestion and differential attachment showed a greater viability with a higher success rate in patch clamp studies.
The sino-atrial node cells can be successfully isolated with enzyme digestion and differential attachment, which requires simple procedures and well preserves the cell viability for better performance in patch clamp experiment.
比较酶消化结合差速贴壁法与酶消化结合差速贴壁加5-溴脱氧尿苷（5-BrdU）（传统）法从新生大鼠分离窦房结细胞用于膜片钳研究的效果。
从新生Wistar大鼠取出窦房结组织，用上述两种方法进行消化并分离成单个窦房结细胞。观察所获得细胞的形态、结构、搏动频率、膜片钳实验的难度及成功率。以酶消化结合差速贴壁法分离的心房细胞作为对照细胞。
培养的窦房细胞中梭形细胞所占比例最大，培养的心房细胞大多呈三角形形态。两种方法分离的窦房细胞大多含有梭形细胞，其中记录到了自发动作电位。酶消化结合差速贴壁法分离的梭形细胞活力更强，在膜片钳研究中的成功率更高。
酶消化结合差速贴壁法可成功分离窦房结细胞，该方法操作简单，能很好地保持细胞活力，在膜片钳实验中表现更佳。
