International Journal of Medical Sciences

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24 September 2017

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Int J Med Sci 2017; 14(9):798-803. doi:10.7150/ijms.19241

Short Research Communication

High Efficiency Low Cost Fibroblast Nucleofection for GMP Compatible Cell-based Gene Therapy

Ziyang Zhang1,2,4,✉, Alex Slobodianski2,3,4, Astrid Arnold4, Jessica Nehlsen4, Ursula Hopfner2, Arndt F. Schilling2,5, Tatjana Perisic2, Hans-Günther Machens2

1. Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
2. Department for Plastic Surgery and Hand Surgery; Klinikum rechts der Isar; Technical University Munich, Munich, Germany;
3. Technical University Munich, Faculty of Medicine, TUM Cells Interdisciplinary Center for Cellular Therapies, Munich, Germany;
4. Department of Plastic Surgery and Hand Surgery, University of Lübeck, Lübeck, Germany;
5. Klinik für Unfallchirurgie, Orthopädie und Plastische Chirurgie, Universitätsmedizin Göttingen, Göttingen, Germany.

Abstract

Background: Dermal fibroblast is a powerful tool for the study of ex vivo DNA delivery in development of both cell therapy and tissue engineering products. Using genetic modification, fibroblasts can be diversely adapted and made suitable for clinical gene therapy. In this study, we first compared several non-viral transfection methods including nucleofection in rat and human primary dermal fibroblast. In addition, the original protocol for nucleofection of primary mammalian fibroblasts was modified in order to achieve the highest possible transfection efficiency, as determined by flow cytometry analysis of the green fluorescent protein (GFP) expression.

Results: the results showed that transfection performance of Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% Fetal Calf Serum (FCS) yielded the best transfection efficiency with rat dermal fibroblasts and ITS (insulin, transferrin, and sodium selenite solution) was comparable to the standard nucleofection solution for human dermal fibroblasts.

Conclusion: Our results suggest a promising application of the modified nucleofection method for GMP compatible therapeutic translational medical research.

Keywords: Dermal fibroblast, nucleofection method, green fluorescent protein

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How to cite this article:
Zhang Z, Slobodianski A, Arnold A, Nehlsen J, Hopfner U, Schilling AF, Perisic T, Machens HG. High Efficiency Low Cost Fibroblast Nucleofection for GMP Compatible Cell-based Gene Therapy. Int J Med Sci 2017; 14(9):798-803. doi:10.7150/ijms.19241. Available from http://www.medsci.org/v14p0798.htm