Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy

Paidikondala, Maruthibabu; Kadekar, Sandeep; Varghese, Oommen P.

doi:10.3390/ijms20010056

Open AccessTechnical Note

Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy

by

Maruthibabu Paidikondala

,

Sandeep Kadekar

and

Oommen P. Varghese

^*

Translational Chemical Biology Laboratory, Polymer Chemistry Division, Department of Chemistry–Ångström Laboratory, Uppsala University, 751 21 Uppsala, Sweden

^*

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2019, 20(1), 56; https://doi.org/10.3390/ijms20010056

Submission received: 18 October 2018 / Revised: 10 December 2018 / Accepted: 20 December 2018 / Published: 23 December 2018

(This article belongs to the Special Issue Biomaterials for Musculoskeletal System)

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Abstract

Ex vivo gene therapy offers enormous potential for cell-based therapies, however, cumbersome in vitro cell culture conditions have limited its use in clinical practice. We have optimized an innovative strategy for the transient transfection of bone morphogenetic protein-2 (BMP-2) expressing plasmids in suspended human stem cells within 5-min that enables efficient loading of the transfected cells into a 3D hydrogel system. Such a short incubation time for lipid-based DNA nanoparticles (lipoplexes) reduces cytotoxicity and at the same time reduces the processing time for cells to be transplanted. The encapsulated human mesenchymal stromal/stem cells (hMSCs) transfected with BMP-2 plasmid demonstrated high expression of an osteogenic transcription factor, namely RUNX2, but not the chondrogenic factor (SOX9), within the first three days. This activation was also reflected in the 7-day and 21-day experiment, which clearly indicated the induction of osteogenesis but not chondrogenesis. We believe our transient transfection method demonstrated in primary MSCs can be adapted for other therapeutic genes for different cell-based therapeutic applications.

Keywords: hydrogel; DNA; transfection; ex vivo; hyaluronic acid

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Paidikondala, M.; Kadekar, S.; Varghese, O.P. Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy. Int. J. Mol. Sci. 2019, 20, 56. https://doi.org/10.3390/ijms20010056

AMA Style

Paidikondala M, Kadekar S, Varghese OP. Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy. International Journal of Molecular Sciences. 2019; 20(1):56. https://doi.org/10.3390/ijms20010056

Chicago/Turabian Style

Paidikondala, Maruthibabu, Sandeep Kadekar, and Oommen P. Varghese. 2019. "Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy" International Journal of Molecular Sciences 20, no. 1: 56. https://doi.org/10.3390/ijms20010056

APA Style

Paidikondala, M., Kadekar, S., & Varghese, O. P. (2019). Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy. International Journal of Molecular Sciences, 20(1), 56. https://doi.org/10.3390/ijms20010056

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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Innovative Strategy for 3D Transfection of Primary Human Stem Cells with BMP-2 Expressing Plasmid DNA: A Clinically Translatable Strategy for Ex Vivo Gene Therapy

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