Astrocytes derived from fetal neural progenitor cells as a novel source for therapeutic adenosine delivery.
OBJECTIVE
Intracerebral delivery of anti-epileptic compounds represents a novel strategy for the treatment of refractory epilepsy. Adenosine is a possible candidate for local delivery based on its proven anti-epileptic effects. Neural stem cells constitute an ideal cell source for intracerebral transplantation and long-term drug delivery. In order to develop a cell-based system for the long-term delivery of adenosine, we isolated neural progenitor cells from adenosine kinase deficient mice (Adk(-/-)) and compared their differentiation potential and adenosine release properties with corresponding wild-type cells.


METHODS
Fetal neural progenitor cells were isolated from the brains of Adk(-/-) and C57BL/6 mice fetuses and expanded in vitro. Before and after neural differentiation, supernatants were collected and assayed for adenosine release using liquid chromatography-tandem mass spectrometry (LC-MS/MS).


RESULTS
Adk(-/-) cells secreted significantly more adenosine compared to wild-type cells at any time point of differentiation. Undifferentiated Adk(-/-) cells secreted 137+/-5 ng adenosine per 10(5) cells during 24 h in culture, compared to 11+/-1 ng released from corresponding wild-type cells. Adenosine release was maintained after differentiation as differentiated Adk(-/-) cells continued to release significantly more adenosine per 24 h (47+/-1 ng per 10(5) cells) compared to wild-type cells (3+/-0.2 ng per 10(5) cells).


CONCLUSIONS
Fetal neural progenitor cells isolated from Adk(-/-) mice--but not those from C57BL/6 mice--release amounts of adenosine considered to be of therapeutic relevance.
Van Dycke
Annelies
A
Laboratory for Clinical and Experimental Neurophysiology, Department of Neurology, Ghent University Hospital, 1K12, 185 De Pintelaan, 9000 Ghent, Belgium. Annelies.VanDycke@UGent.be
Raedt
Robrecht
R
Verstraete
Alain
A
Theofilas
Panos
P
Wadman
Wytse
W
Vonck
Kristl
K
Boison
Detlev
D
Boon
Paul
P
D000241:Adenosine / Q000008:administration & dosage*; D000248:Adenosine Kinase / Q000172:deficiency / Q000235:genetics; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D015153:Blotting, Western; D002454:Cell Differentiation; D002853:Chromatography, Liquid; D004827:Epilepsy / Q000188:drug therapy*; D005260:Female; D053686:Fetal Stem Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D007150:Immunohistochemistry; D007276:Injections, Intraventricular; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016133:Polymerase Chain Reaction; D033581:Stem Cell Transplantation / Q000379:methods*
D000248:Adenosine Kinase; D000241:Adenosine