Design and Validation of a Process Based on Cationic Niosomes for Gene Delivery into Novel Urine-Derived Mesenchymal Stem Cells
Yerai Vado,
Gustavo Puras,
Melania Rosique,
Cesar Martin,
Jose Luis Pedraz,
Shifa Jebari-Benslaiman,
Marian M. de Pancorbo,
Jon Zarate,
Guiomar Perez de Nanclares
Affiliations
Yerai Vado
NanoBioCel Research Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Araba, Spain
Gustavo Puras
NanoBioCel Research Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Araba, Spain
Melania Rosique
BIOMICs Research Group, Microfluidics Cluster UPV/EHU, Lascaray Research Center, University of the Basque Country UPV/EHU, 01009 Vitoria-Gasteiz, Araba, Spain
Cesar Martin
Biofisika Institute (UPV/EHU, CSIC), Department Biochemistry and Molecular Biology, University of the Basque Country University (UPV/EHU), 48940 Leioa, Bizkaia, Spain
Jose Luis Pedraz
NanoBioCel Research Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Araba, Spain
Shifa Jebari-Benslaiman
Biofisika Institute (UPV/EHU, CSIC), Department Biochemistry and Molecular Biology, University of the Basque Country University (UPV/EHU), 48940 Leioa, Bizkaia, Spain
Marian M. de Pancorbo
BIOMICs Research Group, Microfluidics Cluster UPV/EHU, Lascaray Research Center, University of the Basque Country UPV/EHU, 01009 Vitoria-Gasteiz, Araba, Spain
Jon Zarate
NanoBioCel Research Group, Laboratory of Pharmacy and Pharmaceutical Technology, Faculty of Pharmacy, University of the Basque Country UPV/EHU, 01006 Vitoria-Gasteiz, Araba, Spain
Guiomar Perez de Nanclares
Rare Diseases Research Group, Molecular (Epi) Genetics Laboratory, BioAraba Health Research Institute, Araba University Hospital-Txagorritxu, 01009 Vitoria-Gasteiz, Araba, Spain
Background: Mesenchymal stem cells (MSCs) are stem cells present in adult tissues. They can be cultured, have great growth capacity, and can differentiate into several cell types. The isolation of urine-derived mesenchymal stem cells (hUSCs) was recently described. hUSCs present additional benefits in the fact that they can be easily obtained noninvasively. Regarding gene delivery, nonviral vectors based on cationic niosomes have been used and are more stable and have lower immunogenicity than viral vectors. However, their transfection efficiency is low and in need of improvement. Methods: We isolated hUSCs from urine, and the cell culture was tested and characterized. Different cationic niosomes were elaborated using reverse-phase evaporation, and they were physicochemically characterized. Then, they were screened into hUSCs for transfection efficiency, and their internalization was evaluated. Results: GPxT-CQ at a lipid/DNA ratio of 5:1 (w/w) had the best transfection efficiency. Intracellular localization studies confirmed that nioplexes entered mainly via caveolae-mediated endocytosis. Conclusions: In conclusion, we established a protocol for hUSC isolation and their transfection with cationic niosomes, which could have relevant clinical applications such as in gene therapy. This methodology could also be used for creating cellular models for studying and validating pathogenic genetic variants, and even for performing functional studies. Our study increases knowledge about the internalization of tested cationic niosomes in these previously unexplored cells.
