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Article

Efficient Non-Viral Gene Modification of Mesenchymal Stromal Cells from Umbilical Cord Wharton’s Jelly with Polyethylenimine

1
Chemical and Biochemical Processes Research Group, Department of Chemical and Environmental Engineering, Faculty of Engineering, Universidad Nacional de Colombia, Bogotá D.C. 111321, Colombia
2
Advanced Therapies Unit, Instituto Distrital de Ciencia, Biotecnología e Innovación en Salud (IDCBIS), Bogotá D.C. 111611, Colombia
*
Author to whom correspondence should be addressed.
Pharmaceutics 2020, 12(9), 896; https://doi.org/10.3390/pharmaceutics12090896
Received: 4 August 2020 / Revised: 9 September 2020 / Accepted: 10 September 2020 / Published: 22 September 2020
(This article belongs to the Special Issue Gene Delivery Vectors and Physical Methods: Present and Future Trends)
Mesenchymal stromal cells (MSC) derived from human umbilical cord Wharton’s jelly (WJ) have a wide therapeutic potential in cell therapy and tissue engineering because of their multipotential capacity, which can be reinforced through gene therapy in order to modulate specific responses. However, reported methodologies to transfect WJ-MSC using cationic polymers are scarce. Here, WJ-MSC were transfected using 25 kDa branched- polyethylenimine (PEI) and a DNA plasmid encoding GFP. PEI/plasmid complexes were characterized to establish the best transfection efficiencies with lowest toxicity. Expression of MSC-related cell surface markers was evaluated. Likewise, immunomodulatory activity and multipotential capacity of transfected WJ-MSC were assessed by CD2/CD3/CD28-activated peripheral blood mononuclear cells (PBMC) cocultures and osteogenic and adipogenic differentiation assays, respectively. An association between cell number, PEI and DNA content, and transfection efficiency was observed. The highest transfection efficiency (15.3 ± 8.6%) at the lowest toxicity was achieved using 2 ng/μL DNA and 3.6 ng/μL PEI with 45,000 WJ-MSC in a 24-well plate format (200 μL). Under these conditions, there was no significant difference between the expression of MSC-identity markers, inhibitory effect on CD3+ T lymphocytes proliferation and osteogenic/adipogenic differentiation ability of transfected WJ-MSC, as compared with non-transfected cells. These results suggest that the functional properties of WJ-MSC were not altered after optimized transfection with PEI. View Full-Text
Keywords: gene therapy; differentiation; cationic polymer; immunophenotype; immunomodulation; cell therapy; standardization; polyplexes gene therapy; differentiation; cationic polymer; immunophenotype; immunomodulation; cell therapy; standardization; polyplexes
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MDPI and ACS Style

Ramos-Murillo, A.I.; Rodríguez, E.; Beltrán, K.; Ricaurte, C.; Camacho, B.; Salguero, G.; Godoy-Silva, R.D. Efficient Non-Viral Gene Modification of Mesenchymal Stromal Cells from Umbilical Cord Wharton’s Jelly with Polyethylenimine. Pharmaceutics 2020, 12, 896. https://doi.org/10.3390/pharmaceutics12090896

AMA Style

Ramos-Murillo AI, Rodríguez E, Beltrán K, Ricaurte C, Camacho B, Salguero G, Godoy-Silva RD. Efficient Non-Viral Gene Modification of Mesenchymal Stromal Cells from Umbilical Cord Wharton’s Jelly with Polyethylenimine. Pharmaceutics. 2020; 12(9):896. https://doi.org/10.3390/pharmaceutics12090896

Chicago/Turabian Style

Ramos-Murillo, Ana I., Elizabeth Rodríguez, Karl Beltrán, Cristian Ricaurte, Bernardo Camacho, Gustavo Salguero, and Rubén D. Godoy-Silva 2020. "Efficient Non-Viral Gene Modification of Mesenchymal Stromal Cells from Umbilical Cord Wharton’s Jelly with Polyethylenimine" Pharmaceutics 12, no. 9: 896. https://doi.org/10.3390/pharmaceutics12090896

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