Astroglial-Conditioned Media and Growth Factors Modulate Proliferation and Differentiation of Astrocytes in Primary Culture |
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| Authors: | Vincenzo Bramanti Agata Campisi Daniele Tomassoni Antonino Costa Alfredo Fisichella Venera Mazzone Luca Denaro Marcello Avitabile Francesco Amenta Roberto Avola |
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| Affiliation: | (1) Section of Biochemistry and Molecular Biology, Department of Chemical Sciences, University of Catania, Viale Andrea Doria, 6, Catania, Italy;(2) Departments of Biological Chemistry, Medical Chemistry and Molecular Biology, University of Catania, Catania, Italy;(3) Section of Human Anatomy, Department of Experimental Medicine and Public Health, University of Camerino, Camerino, Italy;(4) Section of Urgency and Internal Medicine, Department of Internal Medicine and Systemic Pathologies, University of Catania, Catania, Italy;(5) Section of Human Anatomy, Department “G.F. Ingrassia”, University of Catania, Catania, Italy;(6) Institute of Neurosurgery, Catholic University of Sacred Heart, Rome, Italy |
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| Abstract: | Astroglial conditioned media (ACM) influence the development and maturation of cultured nerve cells and modulate neuron-glia interaction. To clarify mechanisms of astroglial cell proliferation/differentiation in culture, incorporation of [methyl-3H]-thymidine or [5,6-3H]-uridine in cultured astrocytes was assessed. Cultures were pre-treated with epidermal growth factor (EGF), insulin (INS), insulin-like growth factor-I (IGF-I), and basic fibroblast growth factor (bFGF) and subsequently with ACM. DNA labeling revealed a marked stimulatory effect of ACM from 15 days in vitro (DIV) cultures in 30 DIV astrocytes after12 h pre-treatment with growth factors. The main effects were found after INS or EGF pre-treatment in 30 DIV cultures. ACM collected from 15 or 60 or 90 DIV increased RNA labeling of 15 and 30 DIV astrocyte cultures, being the highest value that of 30 DIV cultures added with ACM from 90 DIV. The findings of increased DNA labeling after EGF or INS pre-treatment in 30 DIV cultures, followed by addition of ACM from 15 DIV cultures, suggest that these phenomena may depend by extra cellular signal-regulated kinase 1 (ERK1) activation. |
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| Keywords: | Astroglial conditioned media Growth factors Astrocyte cultures Proliferation DNA and RNA labeling Western blot |
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