Efficient Non-Viral Gene Modification of Mesenchymal Stromal Cells from Umbilical Cord Wharton’s Jelly with Polyethylenimine
Open Access
- 21 September 2020
- journal article
- research article
- Published by MDPI AG in Pharmaceutics
- Vol. 12 (9), 896
- https://doi.org/10.3390/pharmaceutics12090896
Abstract
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.Funding Information
- Universidad Nacional de Colombia (203010026990)
This publication has 64 references indexed in Scilit:
- Wharton’s Jelly-Derived Mesenchymal Stem Cells: Phenotypic Characterization and Optimizing Their Therapeutic Potential for Clinical ApplicationsInternational Journal of Molecular Sciences, 2013
- Stem Cells and Gene Therapy for Cartilage RepairStem Cells International, 2012
- How to screen non-viral gene delivery systems in vitro?Journal of Controlled Release, 2011
- Nonviral Gene Delivery to Mesenchymal Stem Cells Using Cationic Liposomes for Gene and Cell TherapyJournal of Biomedicine and Biotechnology, 2010
- Proliferation and osteoblastic differentiation of bone marrow stem cells: comparison of vertebral body and iliac crestEuropean Spine Journal, 2010
- Fluid Phase Endocytosis Contributes to Transfection of DNA by PEI-25Molecular Therapy, 2009
- The heterogeneous nature of polyethylenimine-DNA complex formation affects transient gene expressionCytotechnology, 2009
- Concise Review: Wharton's Jelly-Derived Cells Are a Primitive Stromal Cell PopulationThe International Journal of Cell Cloning, 2007
- Differential intracellular distribution of DNA complexed with polyethylenimine (PEI) and PEI-polyarginine PTD influences exogenous gene expression within live COS-7 cellsGenetic Vaccines and Therapy, 2007
- Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statementCytotherapy, 2006