HB-EGF affects astrocyte morphology, proliferation, differentiation, and the expression of intermediate filament proteins
Journal article, 2014

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. HB-EGF (heparin-binding EGF-like growth factor) was previously suggested to replace serum, a common and undefined component in primary astrocyte cultures. We show that both in standard 2D and in our newly developed Bioactive3D culture system, HB-EGF affects astrocyte morphology, proliferation, gene/protein expression and leads to partial de-differentiation of astrocytes. Thus, HB-EGF should only be used with caution as a serum replacement in astrocyte cultures.

astrocytes

astrocyte culture

3D cell culture

intermediate filament system

HB-EGF

Author

Till B. Puschmann

University of Gothenburg

Carl Zandén

Chalmers, Applied Physics, Electronics Material and Systems Laboratory

Isabell Lebkuechner

University of Gothenburg

Camille Philippot

University of Gothenburg

Yolanda de Pablo

University of Gothenburg

Johan Liu

Chalmers, Applied Physics, Electronics Material and Systems Laboratory

Milos Pekny

University of Gothenburg

Journal of Neurochemistry

0022-3042 (ISSN) 1471-4159 (eISSN)

Vol. 128 6 878-889

Subject Categories

Biomaterials Science

DOI

10.1111/jnc.12519

PubMed

24188029

More information

Created

10/7/2017