Abstract
Adenosine is an efficient inhibitor of neuronal activity with the ability to suppress seizure activity in various animal models of epilepsy. In the present study adenosine-releasing neuronal cells were generated as a potential source for therapeutically active grafts. Mice with a genetic disruption of the gene encoding adenosine kinase (
Adk
−/−)—the major adenosine metabolizing enzyme—were used as a source for the derivation of adenosine releasing neuronal cells. Since homozygous
Adk
−/− mice constitute a lethal phenotype, embryonic neuroectoderm was derived from intercrosses of
Adk
+/−-mice. Therefore, a rapid genotyping procedure had to be developed using a fluorescent 5′-exonuclease (TaqMan™) assay, which permitted the genotyping of embryonic cell material within 3 h. During this time period the cells to be grafted displayed an unaltered viability. Cultured neuroectodermal
Adk
−/− cells released up to 2 μg adenosine per mg protein per hour.
Adk
−/− neuroectoderm grafted into the lateral brain ventricle of adult mice was found to survive for at least 6 weeks. The method described here suggests the feasibility to graft adenosine releasing neuroectodermal cells as a potential therapeutic approach for the treatment of pharmacoresistant epilepsy.