[A method of primary cell culture of rat pulmonary artery smooth muscle cells and effects of platelet-derived growth factor induced proliferation and migration]

Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2016 Apr 8;32(4):343-346. doi: 10.13459/j.cnki.cjap.2016.04.015.
[Article in Chinese]

Abstract

Objective: To establish an easy, not depending on advanced laboratory apparatus method to isolate and culture rat pulmonary artery smooth muscle cells (PASMCs), and to explore the effects of platelet-derived growth factor (PDGF) on cell proliferation and migration.

Methods: The right ventricle was perfused with the mixture of iron, agarose, and the PASMCs and iron could adhere to agarose. The iron-con-taining tissue would move to side of the tube next to the magnet and could be digested by collagenase I. By the method, vessel-containing tissue could be attained. With 3-4 weeks' purification, the PASMCs could be obtained. The PASMCs morphology was observed by an inverted micro-scope, and identified by immunocytochemistry and immunofluorescence. The effects of PDGF on cell proliferation and migration was detected by MTT assay and scratch wound assay.

Results: 14 days、21 days and primary culture after isolation, the PASMCs was identified, and the re-sult showed that isolation and primary culture of the cells were PASMCs. Compared with the cells with no stimulation, the proliferation of PASMCs exposed to PDGF was increased significantly(P<0.05), and scratch wound assay demonstrated that PDGF induced the significant increase of migration of PASMCs.

Conclusions: This method to isolate and culture rat PASMCs is simple, not depending on advanced laborato-ry. PDGF can promote the proliferation and migration of PASMCs.

Keywords: isolation; primary cell culture; pulmonary arterial smooth muscle cells; rat.

MeSH terms

  • Animals
  • Cell Proliferation
  • Cells, Cultured
  • Muscle, Smooth, Vascular / cytology
  • Myocytes, Smooth Muscle / cytology*
  • Myocytes, Smooth Muscle / drug effects
  • Platelet-Derived Growth Factor / pharmacology*
  • Primary Cell Culture*
  • Pulmonary Artery / cytology
  • Rats

Substances

  • Platelet-Derived Growth Factor