Lentiviral Transduction of Chondrocytes

Lentiviral transduction was performed with chondrocytes encapsulated in alginate beads using lentiviral shRNA vectors expressing EGFP as a reporter marker, as described [8 (link), 10 (link), 41 ]. Viruses were prepared by cotransfecting 293T cells with plasmids coding for the viral components pCMV-VSVG and pCMV-deltaR8-91. After 48 h, viral supernatants were filtered (0.45 μm) and added together with 4 μg/mL polybrene to the chondrocyte cultures. The effectiveness of the transduction was monitored by visualization of the chondrocyte spheres for green fluorescent protein expression (FLOID microscope Life Technologies).

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Mesuraca M., Galasso O., Guido L., Chiarella E., Scicchitano S., Vatrinet R., Morrone G., Bond H.M, & Gasparini G. (2014). Expression Profiling and Functional Implications of a Set of Zinc Finger Proteins, ZNF423, ZNF470, ZNF521, and ZNF780B, in Primary Osteoarthritic Articular Chondrocytes. Mediators of Inflammation, 2014, 318793.

Publication 2014

FLOID microscope

293t cells Alginate Chondrocyte Green fluorescent protein Microscope Plasmids Polybrene Shrna Vectors Viral components Viruses

Corresponding Organization : Magna Graecia University

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Variable analysis

independent variables

Lentiviral transduction of chondrocytes encapsulated in alginate beads using lentiviral shRNA vectors expressing EGFP as a reporter marker

dependent variables

Effectiveness of the transduction monitored by visualization of the chondrocyte spheres for green fluorescent protein expression

control variables

Chondrocytes encapsulated in alginate beads
Lentiviral shRNA vectors expressing EGFP as a reporter marker
Viruses prepared by cotransfecting 293T cells with plasmids coding for the viral components pCMV-VSVG and pCMV-deltaR8-91
Viral supernatants filtered (0.45 μm) and added together with 4 μg/mL polybrene to the chondrocyte cultures

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