High-throughput transfection of interfering RNA into a 3D cell-culture chip

A method for high-throughput retroviral transfection of genes and interfering RNA into 3D cell-culture microarrays is described. 3D cultures more closely mimic the in vivo cellular milieu, thus providing cellular responses to genetic manipulation more similar to the in vivo situation than 2D cultures. This technique is applied to transfect several "toxic" short-hairpin RNAs (shRNAs) into 3D cell cultures. It is demonstrated that the toxicity is similar to that obtained by conventional (non-high-throughput) retroviral transfection of cells grown in similar 3D culture microarrays.