Efficient Nonviral Transfection of Primary Intervertebral Disc Cells by Electroporation for Tissue Engineering Application.

May, Rahel Deborah; Tekari, Adel; Frauchiger, Daniela Angelika; Krismer, A; Benneker, LM; Gantenbein, Benjamin (2017). Efficient Nonviral Transfection of Primary Intervertebral Disc Cells by Electroporation for Tissue Engineering Application. Tissue engineering Part C Methods, 23(1), pp. 30-37. Mary Ann Liebert 10.1089/ten.TEC.2016.0355

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Low back pain (LBP) is an increasing global health problem associated with intervertebral disc (IVD) trauma and degeneration. Current treatment options include surgical interventions with partial unsatisfactory outcomes reported such as failure to relieve LBP, nonunions, nerve injuries, or adjacent segment disease. Cell-based therapy and tissue engineered IVD constructs supplemented with transfected disc cells that incorporate factors enhancing matrix synthesis represent an appealing approach to regenerate the IVD. Gene delivery approaches using transient nonviral gene therapy by electroporation are of a high clinical translational value since the incorporated DNA is lost after few cell generations, leaving the host's genome unmodified. Human primary cells isolated from clinically relevant samples were generally found very hard to transfect compared to cell lines. In this study, we present a range of parameters (voltage pulse, number, and duration) from the Neon(®) Transfection System for efficient transfection of human and bovine IVD cells. To demonstrate efficiency, these primary cells were exemplarily transfected with the commercially available plasmid pCMV6-AC-GFP tagged with copepod turbo green fluorescent protein. Flow cytometry was subsequently applied to quantify transfection efficiency. Our results showed that two pulses of 1400 V for 20 ms revealed good and reproducible results for both human and bovine IVD cells with efficiencies ≥47%. The presented parameters allow for successful human and bovine IVD cell transfection and provide an opportunity for subsequent regenerative medicine application.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > Institute for Surgical Technology & Biomechanics ISTB
04 Faculty of Medicine > Department of Orthopaedic, Plastic and Hand Surgery (DOPH) > Clinic of Orthopaedic Surgery

UniBE Contributor:

May, Rahel Deborah; Tekari, Adel; Frauchiger, Daniela Angelika; Krismer, Anna; Benneker, Lorin Michael and Gantenbein, Benjamin

Subjects:

500 Science > 570 Life sciences; biology
600 Technology > 610 Medicine & health

ISSN:

1937-3384

Publisher:

Mary Ann Liebert

Language:

English

Submitter:

Lilianna Bolliger

Date Deposited:

19 Jul 2017 12:44

Last Modified:

22 Jan 2018 09:07

Publisher DOI:

10.1089/ten.TEC.2016.0355

PubMed ID:

27968705

Uncontrolled Keywords:

DNA delivery; electroporation; intervertebral disc; nonviral gene therapy; tissue engineering applications

BORIS DOI:

10.7892/boris.95564

URI:

https://boris.unibe.ch/id/eprint/95564

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