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.
Institute of Pharmacology and Toxicology, University of Zürich, CH-8057 Zürich, Switzerland.
Denise E
DE
Fedele
Peter
P
Koch
Louis
L
Scheurer
Elizabeth M
EM
Simpson
Hanns
H
Möhler
Oliver
O
Brüstle
Detlev
D
Boison
D000241:Adenosine / Q000378:metabolism*; D000248:Adenosine Kinase / Q000172:deficiency / Q000235:genetics; D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002874:Chromosome Mapping / Q000379:methods; D004622:Embryo, Mammalian; D005455:Fluorescent Antibody Technique / Q000379:methods; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D005818:Genetic Engineering; D005904:Glial Fibrillary Acidic Protein / Q000276:immunology; D006133:Growth Substances / Q000494:pharmacology; D007381:Intermediate Filament Proteins / Q000378:metabolism; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism*; D019081:O Antigens / Q000378:metabolism; D009836:Oligodendroglia / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000502:physiology*
D005904:Glial Fibrillary Acidic Protein; D006133:Growth Substances; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D019081:O Antigens; D000248:Adenosine Kinase; D000241:Adenosine