[Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells].
OBJECTIVE
Set up a method to isolate and identify the small pulmonary arterial smooth muscle cells (PASMCs) in vitro.


METHODS
In sterile conditions, separated the male SD rat pulmonary artery, digested by collagenase I and cultured primary PASMCs. Measured cell viability; observed by phase contrast microscope; identified by immunocytochemistry and immunofluorescence staining as a label for smooth muscle alpha-actin.


RESULTS
PASMCs were identified by morphology and immunocytochemistry, immunofluorescence staining, with the cell viability is over 96.5%. The primary culture could be subcultured after 4-7 days and successfully passaged without change in morphology and growth characteristic.


CONCLUSIONS
This technique has advantage of the method is simple, short cultivate, good reproducibility, the primary cultured PASMCs quantity and the rapid growth.
Qian
Guo-qing
GQ
Department of Respiratory, The First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China.
Wang
Liang-xing
LX
Cheng
Chan
C
Huang
Xiao-ying
XY
Yei
Shu-ting
ST
D000818:Animals; D001160:Arterioles / Q000166:cytology; D002469:Cell Separation / Q000379:methods; D008168:Lung / Q000098:blood supply; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D061251:Primary Cell Culture / Q000379:methods*; D011651:Pulmonary Artery / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley