Journal
JOURNAL OF NEUROCHEMISTRY
Volume 128, Issue 6, Pages 878-889Publisher
WILEY-BLACKWELL
DOI: 10.1111/jnc.12519
Keywords
3D cell culture; astrocyte culture; astrocytes; HB-EGF; intermediate filament system
Categories
Funding
- Swedish Medical Research Council [11548]
- ALF Gothenburg [11392]
- AFA Research Foundation
- Soderbergs Foundations
- Sten A. Olsson Foundation for Research and Culture
- Hjarnfonden
- Hagstromer's Foundation Millennium
- Amlov's Foundation
- E. Jacobson's Donation Fund
- VINNOVA Health Program
- Swedish Stroke Foundation
- Swedish Society of Medicine
- Free Mason Foundation
- Chalmers University of Technology
- NanoNet COST Action [BM1002]
- EU [237956, 279017]
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Heparin-binding epidermal growth factor-like growth factor (HB-EGF), a vascular-derived trophic factor, belongs to the epidermal growth factor (EGF) family of neuroprotective, hypoxia-inducible proteins released by astrocytes in CNS injuries. It was suggested that HB-EGF can replace fetal calf serum (FCS) in astrocyte cultures. We previously demonstrated that in contrast to standard 2D cell culture systems, Bioactive3D culture system, when used with FCS, minimizes the baseline activation of astrocytes and preserves their complex morphology. Here, we show that HB-EGF induced EGF receptor (EGFR) activation by Y1068 phosphorylation, Mapk/Erk pathway activation, and led to an increase in cell proliferation, more prominent in Bioactive3D than in 2D cultures. HB-EGF changed morphology of 2D and Bioactive3D cultured astrocytes toward a radial glia-like phenotype and induced the expression of intermediate filament and progenitor cell marker protein nestin. Glial fibrillary acidic protein (GFAP) and vimentin protein expression was unaffected. RT-qPCR analysis demonstrated that HB-EGF affected the expression of Notch signaling pathway genes, implying a role for the Notch signaling in HB-EGF-mediated astrocyte response. HB-EGF can be used as a FCS replacement for astrocyte expansion and in vitro experimentation both in 2D and Bioactive3D culture systems; however, caution should be exercised since it appears to induce partial de-differentiation of astrocytes.
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