Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells

Sara Palchetti, Daniela Pozzi, Cristina Marchini, Augusto Amici, Cristina Andreani, Caterina Bartolacci, Luca Digiacomo, Valentina Gambini, Francesco Cardarelli, Carmine Di Rienzo, Giovanna Peruzzi, Heinz Amenitsch, Rocco Palermo, Isabella Screpanti, Giulio Caracciolo

Research output: Contribution to journalArticleResearchpeer-review

Abstract

To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine – the gold standard among transfection reagents – was used as a reference. On the basis of our results, we suggest that an additional transfection barrier impairs transfection efficiency, that is: low lipoplex concentration at the cell surface. Based on the acquired knowledge we propose an optimized transfection protocol that allowed us to efficiently transfect DND41, JURKAT, MOLT3, P12-ICHIKAWA, ALL-SILL, TALL-1 human T-cell acute lymphoblastic leukemia (T-ALL) cell lines known to be difficult-to-transfect by using non-viral vectors and where LFN-based technologies fail to give satisfactory results.

Original languageEnglish
Pages (from-to)681-691
Number of pages11
JournalNanomedicine / Nanotechnology, Biology, and Medicine
Volume13
Issue number2
DOIs
Publication statusPublished - 1 Feb 2017

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Keywords

  • Cationic liposomes
  • Cell transfection
  • Hard-to-transfect cells
  • Lipoplexes
  • Transfection efficiency

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Bioengineering
  • Biomedical Engineering
  • Molecular Medicine
  • Materials Science(all)
  • Pharmaceutical Science

Fields of Expertise

  • Advanced Materials Science
  • Human- & Biotechnology

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