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Cytotechnology

, Volume 69, Issue 5, pp 831–840 | Cite as

Isolation, culture and identification of pulmonary arterial smooth muscle cells from rat distal pulmonary arteries

  • Gongyong PengEmail author
  • Juan Xu
  • Rongmin Liu
  • Zhenli Fu
  • Shaoxing Li
  • Wei Hong
  • Jinglong Chen
  • Bing Li
  • Pixin RanEmail author
Method in Cell Science

Abstract

The culture of pulmonary arterial smooth muscle cells (PASMCs) is one of the most powerful tools for exploring the mechanisms of pulmonary hypertension (PH). Both pulmonary vasoconstriction and remodeling occur predominantly in distal pulmonary arteries (PA). In this study, we provide our detailed and standardized protocol for easy isolation and culture of PASMCs from rat distal PA to supply every investigator with a simple, economical and useful method in studying PH. The protocol can be divided into four stages: isolation of distal PA, isolation of cells, growth in culture and passage of cells. Rat distal PASMCs were characterized by morphological activity and by immunostaining for smooth muscle α-actin and smooth muscle myosin heavy chain, but not for CD90/Thy-1 or von Willebrand factor. Furthermore, functional assessments were performed, confirming the presence of voltage-dependent Ca2+ channels and physiological characteristic of response to hypoxia. In conclusion, we have developed a detailed and simple protocol for obtaining rat distal PASMCs. These PASMCs exhibit features consistent with vascular smooth muscle cells, and they could subsequently be used to further explore the pathophysiological mechanisms of PH.

Keywords

Cell culture Smooth muscle cells Pulmonary artery Rat 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (81570045, 81000020), the Natural Science Foundation of Guangdong Province (2014A030313486), the Scientific Research Grant of Guangzhou City (201510010226), Guangzhou Department of Education Yangcheng Scholarship (10A025G) and the grant of State Key Laboratory of Respiratory Disease (SKLRD2016ZJ013, SKLRD2011QN011).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

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Copyright information

© Springer Science+Business Media Dordrecht 2017

Authors and Affiliations

  • Gongyong Peng
    • 1
    Email author
  • Juan Xu
    • 1
  • Rongmin Liu
    • 1
  • Zhenli Fu
    • 1
  • Shaoxing Li
    • 1
    • 2
  • Wei Hong
    • 3
  • Jinglong Chen
    • 1
  • Bing Li
    • 3
  • Pixin Ran
    • 1
    Email author
  1. 1.Guangzhou Institute of Respiratory Disease, State Key Laboratory of Respiratory DiseaseThe First Affiliated Hospital, Guangzhou Medical UniversityGuangzhouPeople’s Republic of China
  2. 2.Intensive Care UnitGuangzhou Panyu Central HospitalGuangzhouPeople’s Republic of China
  3. 3.The Research Center of Experiment MedicineGuangzhou Medical UniversityGuangzhouPeople’s Republic of China

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