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study Limitations
Cell engraftment and survival was not quantified. Bromodeoxyuridine (BrdU)
or other cellular markers were not used to avoid interference with cell capacity to
colonize and proliferate. Such interference could cause a decrease in the proposed
EPO effect.
Retroviral transduction itself can potentially alter the phenotype of the trans-
duced cells. However, the expression profile of transduced cells did not exhibit
any negative effects when the transduced cell culture supernants were added to
cardiomyocyte culture in vitro.
In order to avoid possible systemic interference, we did not apply an immu-
nosuppression protocol. However, EPO detection 1 month after cell transplanta-
tion suggests that immune response, if evoked, is limited in magnitude. The EPO
tissue level could not be quantified due to technical constraints for detection of
predicted low levels of the protein.
conclusion
In situ expression of rhEPO by genetically engineered cardiac fibroblasts exerts
cytoprotective and pro-angiogenic effects. However, these favorable effects were
not translated into functional improvement after MI. The results suggest that
for successful local EPO-based therapy, several improvements should be made to
determine the optimal cell type for gene delivery, the most effective recombinant
viral vector, myocardial EPO response and, notably, an accurate therapeutic win-
dow for cell and gene transfer.
competing interests
The authors declare that they have no competing interests.
Authors' contributions
ER carried out the in vitro and in vivo experiments, including data acquisition
and analysis. OS participated in study design and coordination, and helped in
drafting the manuscript. AN participated in conceptual study design and coor-
dination, and helped to draft the manuscript. TE provided help in study design,
data acquisition and analysis. MF participated in data acquisition and analysis.
RH carried out the in vivo experiments. AD participated in conceptual study
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