2.4 Cell culture
VSMCs were prepared from thoracic aorta of 50-day-old female
Sprague-Dawley rats. Aortic VSMCs were cultured in Dulbecco’s modified
Eagle’s medium (Gibco, USA) with 10% fetal bovine serum (BI, USA) and
1% penicillin/streptomycin at 37 °C in a 5% CO2humidified incubator. Immunohistochemical staining revealed positivity
for smooth muscle α-actin (Wanleibio, China) and negativity for CD105
(Wanleibio, China), indicating that primary VSMCs of rats were
successfully isolated. Cells in passages 3-9 were used for all
experiments.