Engineering embryonic stem cell derived glia for adenosine delivery

Based on the anticonvulsant and neuroprotective properties of adenosine, and based on the long-term survival potential of stem cell derived brain implants, adenosine releasing stem cells may constitute a novel tool for the treatment of epilepsy. Pluripotency and unlimited self-renewal make embryonic stem (ES) cells a particularly versatile donor source for cell transplantation. With the aim to test the feasibility of a stem cell-based delivery system for adenosine, both alleles of adenosine kinase (ADK), the major adenosine-metabolizing enzyme, were disrupted by homologous recombination in ES cells. Adk ^-/- ES cells were subjected to a glial differentiation protocol and, as a result, gave rise to proliferating glial precursors, which could be further differentiated into mature astrocytes and oligodendrocytes. Thus, a lack of ADK does not compromise the glial differentiation potential of ES cells. The Adk ^-/- ES cells yielded glial populations with an adenosine release of up to 40.1 ± 6.0 ng per 10 ⁵ cells per hour, an amount considered to be sufficient for seizure suppression. Our findings indicate that Adk ^-/- ES cells constitute a potential source for therapeutic adenosine releasing grafts.