, Volume 70, Issue 2, pp 751–760 | Cite as

Primary culture of lung fibroblasts from hyperoxia-exposed rats and a proliferative characteristics study

  • Shi-meng Zhao
  • Hong-min Wu
  • Mei-ling Cao
  • Dan Han
Original Article


Lung fibrosis is an ultimate consequence of bronchopulmonary dysplasia (BPD) which shows the excessive proliferation of lung fibroblasts (LFs). To find a better model for studying the role of LFs in hyperoxia-induced lung fibrosis at the cellular level, we isolated LFs from the lung tissue of hyperoxia- and normoxia-exposed rat lungs on postnatal days 7, 14 and 21 for primary culture to study their proliferative behavior. In the present study, the LF predominance was > 95% in our culture method. The LFs isolated from rats exposed to hyperoxia in vivo showed significantly greater proliferation than that from normoxia-exposed rats. Flow cytometry revealed that percentage of LFs in S and G2/M stage increased, and proportion in the G0/G1 stage declined at the same time. A greater presence of myofibroblasts in LFs isolated from rats exposed to hyperoxia compared with those exposed to normoxia. In addition, elevated collagen level, transforming growth factor-β and connective tissue growth factor protein expression in conditioned medium were also found in hyperoxia LFs. These data demonstrate that hyperoxia promotes LFs proliferation, myofibroblast transdifferentiation and collagen synthesis in a time-dependent manner. The primary culture of LFs from hyperoxia-exposed rats is a feasible method for studying the pathogenesis and treatment of lung fibrosis caused by BPD at the cellular level.


Hyperoxia Lung fibroblast Primary cell culture Proliferative characteristic 



Bronchopulmonary dysplasia


Lung fibroblasts


Transforming growth factor-β


Connective tissue growth factor


Phosphate-buffered saline


Radial alveolar count


The mean septal wall thickness


Fetal bovine serum


Minimum essential medium


Non-fat dry milk


Proliferating cell nuclear antigen


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Shi-meng Zhao
    • 1
  • Hong-min Wu
    • 1
  • Mei-ling Cao
    • 1
  • Dan Han
    • 1
  1. 1.Department of NeonatologyThe First Affiliated Hospital of China Medical UniversityShenyangChina

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