We developed and characterized a 3D collagen hydrogel model for B16.F10 melanoma tumors. Cells inthis 3D environment exhibited lower proliferation than cells in the conventional 2D culture ...environment.Interestingly, the basal expression levels of several genes varied when compared to conventionally growncells. In each growth environment, a significant number of melanoma cells were transfected by plasmidelectroporation (electrotransfer), although expression could only be ascertained on the surface of the 3Dconstructs. Cellular responses to plasmid entry as demonstrated by pro-inflammatory cytokine and che-mokine upregulation varied based on the growth environment, as did the mRNA levels of several putativeDNA-specific pattern recognition receptors (DNA sensors). Unexpectedly, when plasmid DNA was deliv-ered while cells where attached in the 2D or 3D environments, the mRNAs of the DNA sensor p204 andthe inflammatory mediator TNFawere regulated in cells receiving pulses only. However, we were unableto confirm coordinate upregulation of TNFaand p204 proteins. This study confirms that cell responsesdiffer significantly based on their environment, and demonstrates the difficulty of extending experimen-tal observations between cell environments.
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