Insulin and heparin-binding epidermal growth factor-like growth factor synergistically promote astrocyte survival and proliferation in serum-free medium

Mei Jia, Zhongfang Shi, Xu Yan, Lixin Xu, Liping Dong, Jiaxin Li, Yujiao Wang, Shaohua Yang, Fang Yuan

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Background: In vitro systems allowing maintenance and experimentation on primary astrocyte cultures have been used for decades. Astrocyte cultures are most maintained in serum-containing medium which has been found to alter the morphology and gene profiles of astrocytes. New method: Here, we reported a new serum-free medium for astrocyte culture, which consisted of DMEM and NB media supplemented with insulin and heparin-binding epidermal growth factor-like growth factor (HB-EGF) (SF-I-H medium). Meanwhile FBS-containing (FBS) medium composed of DMEM medium containing 10% FBS were used for comparison study. Cerebral cortex was harvested from postnatal day 1 Wistar rats and brain cells were isolated and seeded to poly-L-lysine coated culture dishes after 15 min differential velocity adherence. Results: Compared with FBS medium, astrocytes in SF-I-H medium were smaller and exhibited process bearing morphologies. MTT assays showed that cell density and proliferation rate were higher in SF-I-H medium than in FBS medium all the time, and flow cytometry analysis revealed that SF-I-H medium promoted cell mitosis in a manner comparable to FBS medium. Consistently, western blot analysis further revealed that insulin and HB-EGF synergistically activated the PI3K/AKT and MAPK/ERK1/2 signaling cascades as FBS. Comparison with existing method(s): Astrocytes cultured in SF-I-H medium grew faster than FBS medium. Conclusion: Taken together, our results indicated that SF-I-H medium, in which cell morphology was similar with astrocytes in brain, was more effective for astrocyte survival and proliferation than FBS medium, providing a new cell model to study astrocyte functions without the interference of serum.

Original languageEnglish
Pages (from-to)240-247
Number of pages8
JournalJournal of Neuroscience Methods
Volume307
DOIs
StatePublished - 1 Sep 2018

Keywords

  • Astrocytes
  • Cell proliferation
  • Cell survival
  • Heparin-binding epidermal growth factor-like growth factor
  • Insulin
  • Serum-free medium

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