Back
Acta Physiologica 2009; Volume 195, Supplement 670
Belgian Society for Fundamental and Clinical Physiology and Pharmacology, Spring Meeting 2009
3/7/2009-3/7/2009
Ghent University, Gent, Belgium
ADENOSINE SECRETION BY NEURAL STEM CELLS ISOLATED FROM ADK-/-MICE
Abstract number: P-13
Van Dycke1 A., Raedt1 R., Boison2 D., Verstraete3 A., Meurs1 A., Wyckhuys1 T., El Tahry1 R., Vonck1 K., Wadman1,4 W., Boon1 P.
1Laboratory for Clinical and Experimental Neurophysiology, Department of Neurology, Ghent University Hospital, Ghent, Belgium
2Robert Stone Dow Neurobiology Laboratories, Legacy Research, Portland, USA
3Laboratory of Clinical Biology, Ghent University Hospital, Ghent, Belgium
4Swammerdam Institute of Life Sciences, Department of Neurobiology, University of Amsterdam, Amsterdam, The Netherlands
Purpose:
There is an ongoing search for alternative treatment options for patients with refractory epilepsy. Local intracerebral delivery of anti-epileptic compounds may represent a novel strategy with specific advantages such as the option of higher local doses and reduced side effects. Adenosine is a good candidate for local delivery since it has proven anti-epileptic effects. Neural stem cells isolated from adenosine kinase deficient (Adk-/-) mice overexpress adenosine and may be a source for long-term local delivery. We evaluated the amount of adenosine secretion compared to control cells isolated from C57 Black 6 mice (C57BL/6).
Methods:
Foetal neural stem cells were isolated from Adk-/- and C57BL/6 p14 mice foetuses. Stem cells were cultured and expanded in vitro for several weeks. Then cells were plated at different densities (10.000 and 100.000 cells/cm²) during 24 hours. Medium was replaced and samples (200ml) were taken at different time points (after 1 h, 2h, 4h, 8h, 12h and 24h). The same was done with cells plated at a density of 50.000/cm² that were allowed one week of differentiation. The different samples were analyzed to measure the amount of adenosine release with Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS).
Results:
Adk-/- cells secreted more adenosine compared to control cells at any time point. After 24h 507ng per 100.000 cells was secreted by Adk-/- cells compared to 40ng per 100.000 cells by control cells. The Adk-/- cells plated at a density of 10.000 cells/cm² secreted 10 times less compared to the cells plated at 100.000/cm². Differentiated cells released less adenosine: 89ng (Adk-/-) per 50.000 cells after 24h, but this was also higher compared to the control cells.
Conclusions:
Foetal neural stem cells isolated from Adk-/- mice release significantly more adenosine compared to control cells isolated from C57BL/6 mice. Compared to studies concerning local delivery of adenosine, both non-differentiated and differentiated Adk-/- cells release enough adenosine in vitro per 24h to be a tool for long-term local intracerebral adenosine delivery as a therapy for refractory epilepsy.
To cite this abstract, please use the following information:
Acta Physiologica 2009; Volume 195, Supplement 670 :P-13