TY - JOUR
T1 - Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells
AU - Palchetti, Sara
AU - Pozzi, Daniela
AU - Marchini, Cristina
AU - Amici, Augusto
AU - Andreani, Cristina
AU - Bartolacci, Caterina
AU - Digiacomo, Luca
AU - Gambini, Valentina
AU - Cardarelli, Francesco
AU - Di Rienzo, Carmine
AU - Peruzzi, Giovanna
AU - Amenitsch, Heinz
AU - Palermo, Rocco
AU - Screpanti, Isabella
AU - Caracciolo, Giulio
PY - 2017/2/1
Y1 - 2017/2/1
N2 - 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.
AB - 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.
KW - Cationic liposomes
KW - Cell transfection
KW - Hard-to-transfect cells
KW - Lipoplexes
KW - Transfection efficiency
UR - http://www.scopus.com/inward/record.url?scp=85010451201&partnerID=8YFLogxK
U2 - 10.1016/j.nano.2016.08.019
DO - 10.1016/j.nano.2016.08.019
M3 - Article
AN - SCOPUS:85010451201
SN - 1549-9634
VL - 13
SP - 681
EP - 691
JO - Nanomedicine: Nanotechnology, Biology, and Medicine
JF - Nanomedicine: Nanotechnology, Biology, and Medicine
IS - 2
ER -