Preliminary study on non-viral transfection of F9 (factor IX) gene by nucleofection in human adipose-derived mesenchymal stem cells
Entity
UAM. Departamento de Cirugía; Instituto de Investigación Sanitaria Fundación Jiménez Díaz (IIS-FJD)Publisher
PeerJ, LtdDate
2016-01-01Citation
10.7717/peerj.1907
PeerJ 4 (2016): e1907
ISSN
2167-8359DOI
10.7717/peerj.1907Funded by
This study was supported by the Andalusian Association of Hemophilia (Asanhemo), the Victoria Eugenia Royal Foundation, Octapharma S.A., Baxter Spain and Pfizer SpainEditor's Version
http://dx.doi.org/10.7717/peerj.1907Subjects
Advanced therapies; Factor IX; Gene therapy; Hemophilia B; Human mesenchymal adipose-derived stem cells; Non-viral; Nucleofection; MedicinaRights
© 2016 Olmedillas López et alAbstract
Background. Hemophilia is a rare recessive X-linked disease characterized by a deficiency of coagulation factor VIII or factor IX. Its current treatment is merely palliative. Advanced therapies are likely to become the treatment of choice for the disease as they could provide a curative treatment. Methods. The present study looks into the use of a safe non-viral transfection method based on nucleofection to express and secrete human clotting factor IX (hFIX) where human adipose tissue derived mesenchymal stem cells were used as target cells in vitro studies and NOD. Cg-Prkdcscid Il2rgtm1Wjl/SzJ mice were used to analyze factor IX expression in vivo studies. Previously, acute liver injury was induced by an injected intraperitoneal dose of 500 mg/kg body weight of acetaminophen. Results. Nucleofection showed a percentage of positive cells ranging between 30.7% and 41.9% and a cell viability rate of 29.8%, and cells were shown to secrete amounts of hFIX between 36.8 and 71.9 ng/mL. hFIX levels in the blood of NSG mice injected with ASCs transfected with this vector, were 2.7 ng/mL 48 h after injection. Expression and secretion of hFIX were achieved both in vitro cell culture media and in vivo in the plasma of mice treated with the transfected ASCs. Such cells are capable of eventually migrating to a previously damaged target tissue (the liver) where they secrete hFIX, releasing it to the bloodstream over a period of at least five days from administration. Conclusions. The results obtained in the present study may form a preliminary basis for the establishment of a future ex vivo non-viral gene/cellular safe therapy protocol that may eventually contribute to advancing the treatment of hemophilia.
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Google Scholar:Olmedillas López, Susana
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García Arranz, Mariano Andrés
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García Olmo, Damián
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Liras, Antonio
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