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Robust, Efficient, and Practical Electrogene Transfer Method for Human Mesenchymal Stem Cells Using Square Electric Pulses

Lookup NU author(s): Dr Aaron Liew

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Abstract

Mesenchymal stem cells (MSCs) are multipotent nonhematopoietic cells with the ability to differentiate into various specific cell types, thus holding great promise for regenerative medicine. Early clinical trials have proven that MSC-based therapy is safe, with possible efficacy in various diseased states. Moreover, genetic modification of MSCs to improve their function can be safely achieved using electrogene transfer. We previously achieved transfection efficiencies of up to 32% with preserved viability in rat MSCs. In this study, we further improved the transfection efficiency and transgene expression in human MSCs (hMSCs), while preserving the cells viability and ability to differentiate into osteoblasts and adipocytes by increasing the plasmid concentration and altering the osmotic pressure of the electrotransfer buffer. Using a square-wave electric pulse generator, we achieved a transfection efficiency of more than 80%, with around 70% viability and a detectable transgene expression of up to 30 days. Moreover, we demonstrated that this transfection efficiency can be reproduced reliably on two different sources of hMSCs: the bone marrow and adipose tissue. We also showed that there was no significant donor variability in terms of their transfection efficiency and viability. The cell confluency before electrotransfer had no significant effect on the transfection efficiency and viability. Cryopreservation of transfected cells maintained their transgene expression and viability upon thawing. In summary, we are reporting a robust, safe, and efficient protocol of electrotransfer for hMSCs with several practical suggestions for an optimal use of genetically engineered hMSCs for clinical application.

Publication metadata

Author(s): Liew A, André FM, Lesueur LL, DeMénorval MA, O'Brien T, Mir LM

Publication type: Article

Publication status: Published

Journal: Human Gene Therapy Methods

Year: 2013

Volume: 24

Issue: 5

Pages: 289-297

Print publication date: 30/08/2013

ISSN (print): 1946-6536

ISSN (electronic): 1946-6544

Publisher: Mary Ann Liebert, Inc. Publishers

URL: http://dx.doi.org/10.1089/hgtb.2012.159

DOI: 10.1089/hgtb.2012.159

PubMed id: 23931158

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