Erythropoietin producing cells for potential cell therapy.
BACKGROUND
Anemia is an inevitable outcome of chronic renal failure due to the kidney's decreased ability to produce erythropoietin (EPO). We examined the feasibility of isolating and expanding EPO-producing cells for cell-based therapy.


METHODS
Renal cells from 7- to 10-day-old mice were culture-expanded. The cells at each subculture stage were characterized for EPO expression, using immunocytochemistry, FACS, and Western Blot analysis, with EPO-specific antibodies. To assess the levels of EPO expression, cells incubated under normoxic and hypoxic conditions were analyzed.


RESULTS
Immunocytochemical analysis of the cultured renal cells expressed EPO at each subculture stage (P1-P3). Western Blot analysis of the detergent-solubilized cell extracts detected EPO (34 kDa) protein in the kidney cells of all passages tested.


CONCLUSIONS
These results demonstrate that EPO-producing renal cells can be grown and expanded in culture. The cells stably expressed EPO at multiple subculture stages and they are able to form tissue in vivo. This study shows that EPO-producing cells may be used as a potential treatment option for anemia caused by chronic renal failure.
Aboushwareb
Tamer
T
Wake Forest Institute for Regenerative Medicine and Department of Urology, Wake Forest University Health Sciences, Medical Center Boulevard, Winston-Salem, NC 27157, USA.
Egydio
Fernanda
F
Straker
Lauren
L
Gyabaah
Kenneth
K
Atala
Anthony
A
Yoo
James J
JJ
D000740:Anemia / Q000209:etiology / Q000628:therapy*; D000818:Animals; D017690:Cell Transplantation; D002478:Cells, Cultured; D004921:Erythropoietin / Q000378:metabolism*; D005240:Feasibility Studies; D005434:Flow Cytometry; D016130:Immunophenotyping; D007672:Kidney Cortex / Q000166:cytology*; D007676:Kidney Failure, Chronic / Q000150:complications*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL
D004921:Erythropoietin