title: [A method for isolating and identifying newborn rat sino-atrial node cells for patch-clamp study].
abstract: OBJECTIVE
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Cardiology, Guang'anmen Hospital, China Academy of Chinese Medical Sciences, Beijing 100053, China. wangyanlihappy@yahoo.com.cn
authors.forename: Yanli
authors.initials: Y
authors.lastname: Wang
authors.forename: Ruxiu
authors.initials: R
authors.lastname: Liu
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D018929:Cell Culture Techniques; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar; D012849:Sinoatrial Node / Q000166:cytology*